			SEQUENCE LISTING

<110> CODEXIS, INC.
 
<120> PENICILLIN-G ACYLASES
<130> CX2-163USP1
 
<140> TO BE ASSIGNED
<141> SUBMITTED HEREWITH
 
 
<160> 90
 
<210> 1
<211> 2541
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Penicillin G Acylase From Kluyvera Citrophila
 
<400> 1
atgaaaaata gaaatcgtat gatcgtgaac ggtattgtga cttccctgat ctgttgttct 60
agcctgtcag cgctggcggc aagcccgcca accgaggtta agatcgttcg cgatgaatac 120
ggcatgccgc atatttacgc cgatgatacc tatcgactgt tttacggcta tggctacgtg 180
gtggcgcagg atcgcctgtt ccagatggaa atggcgcgcc gcagtactca ggggaccgtc 240
tccgaggtgc tgggcaaagc attcgtcagt tttgataaag atattcgcca gaactactgg 300
ccggattcta ttcgcgcgca gatagcttcc ctctccgctg aggataaatc cattctgcag 360
ggctatgccg atggcatgaa tgcgtggatc gataaagtga acgccagccc cgataagctg 420
ttaccccagc agttctccac ctttggtttt aaacccaagc attgggaacc gtttgatgtg 480
gcgatgattt ttgtcggcac catggcgaac cggttttctg acagcaccag cgaaattgat 540
aacctggcgc tgctgacggc gctaaaagat aaatacggca agcagcaggg catggcggtc 600
tttaaccagc tgaaatggct ggttaatcct tccgcgccaa ccaccattgc ggcgcgggaa 660
agcgcctatc cgctgaagtt tgatctgcaa aacacgcaaa cggcggcgct gctgccgcgc 720
tacgaccagc cggcaccgat gctcgaccgc ccggcaaaag ggaccgatgg cgcgctgctg 780
gcgctgaccg ccgatcagaa ccgggaaact atcgccgcgc agttcgcgca aagcggcgct 840
aacggcctgg ctggctaccc gaccactagc aatatgtggg tgattggcaa aaacaaagcc 900
caggatgcga aggccattat ggtcaatggg ccgcagtttg gttggtatgc gccggcgtac 960
acctacggta tcggcctgca cggcgcgggc tatgacgtca ccggcaatac gccgtttgcc 1020
tatccgggcc tcgtttttgg tcacaacggc accatttcat ggggatccac cgccggtttt 1080
ggtgatgatg tcgatatctt tgccgaaaaa ctttccgccg agaagccggg ctattaccag 1140
cataacggcg agtgggtgaa gatgttgagc cgcaaggaga ctattgcggt caaagacggc 1200
cagccggaga cctttaccgt ttggcgcacg ctgcacggca acgtcattaa aaccgatact 1260
gcgacgcaga ccgcctatgc caaagcgcgc gcctgggatg gcaaagaggt ggcgtccctg 1320
ctggcgtgga cgcaccagat gaaggccaaa aactggccgg agtggacgca gcaggcggcc 1380
aaacaggcgc tgaccattaa ctggtactac gccgatgtga acggcaatat cggctatgtg 1440
cataccggcg cctatccgga tcgccagccc ggccacgacc cgcgtttgcc ggttcccggc 1500
actggaaaat gggactggaa agggttgctg tcgtttgatt tgaatccgaa agtgtataac 1560
ccgcagtcgg gctatatcgc caactggaac aactcgccgc aaaaagacta cccggcctct 1620
gatctgttcg cgttcctgtg gggcggtgcg gatcgagtta ctgagatcga cacgatcctc 1680
gataagcaac cgcgcttcac cgccgatcag gcgtgggatg tgatccgcca aaccagccgt 1740
cgggatctca acctgcggtt gttcttaccg gcgctgaagg acgccaccgc gaacctggcg 1800
gaaaacgatc cgcgccgcca actggtggat aaactggcga gctgggacgg tgaaaacctt 1860
gtcaacgatg acggaaaaac ctatcagcaa ccgggatcgg cgattcttaa cgcctggctg 1920
accagcatgc tcaagcgcac ggtggttgcc gcggtcccag cgccgtttgg caagtggtac 1980
agcgccagtg gctatgaaac cacccaggac gggccaaccg gctcgctgaa catcagcgtg 2040
ggggcgaaaa tcctctacga agctctgcag ggtgataagt cgccaatccc gcaggcggtc 2100
gatctgtttg gcgggaaacc gcagcaggaa gtgatactgg cggcgctgga cgacgcttgg 2160
cagacgctgt caaaacgcta cggtaacgac gtcaccggct ggaaaacccc tgccatggcg 2220
cttaccttcc gggccaataa cttcttcggc gtgccgcagg cggcagcaaa agaggcgcgt 2280
catcaggcgg agtaccagaa ccgcggtacg gaaaacgaca tgattgtctt ctcaccgacg 2340
tcgggtaacc gcccggttct tgcctgggat gtggtggcgc cggggcaaag cggttttatc 2400
gcgccggatg gcaaagccga taagcactat gacgatcagc tgaaaatgta cgagagcttt 2460
ggccgtaaat cgctgtggtt aacgcctcag gacgttgacg agcacaaaga gtctcaggaa 2520
gtgctgcagg tacagcgcta a                                           2541

 
<210> 2
<211> 846
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 2
Met Lys Asn Arg Asn Arg Met Ile Val Asn Gly Ile Val Thr Ser Leu 
1               5                   10                  15      
Ile Cys Cys Ser Ser Leu Ser Ala Leu Ala Ala Ser Pro Pro Thr Glu 
            20                  25                  30          
Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala Asp 
        35                  40                  45              
Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln Asp 
    50                  55                  60                  
Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr Val 
65                  70                  75                  80  
Ser Glu Val Leu Gly Lys Ala Phe Val Ser Phe Asp Lys Asp Ile Arg 
                85                  90                  95      
Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu Ser 
            100                 105                 110         
Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn Ala 
        115                 120                 125             
Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln Gln 
    130                 135                 140                 
Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp Val 
145                 150                 155                 160 
Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser Thr 
                165                 170                 175     
Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys Tyr 
            180                 185                 190         
Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu Val 
        195                 200                 205             
Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr Pro 
    210                 215                 220                 
Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala Ala Leu Leu Pro Arg 
225                 230                 235                 240 
Tyr Asp Gln Pro Ala Pro Met Leu Asp Arg Pro Ala Lys Gly Thr Asp 
                245                 250                 255     
Gly Ala Leu Leu Ala Leu Thr Ala Asp Gln Asn Arg Glu Thr Ile Ala 
            260                 265                 270         
Ala Gln Phe Ala Gln Ser Gly Ala Asn Gly Leu Ala Gly Tyr Pro Thr 
        275                 280                 285             
Thr Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys 
    290                 295                 300                 
Ala Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Ala Pro Ala Tyr 
305                 310                 315                 320 
Thr Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn 
                325                 330                 335     
Thr Pro Phe Ala Tyr Pro Gly Leu Val Phe Gly His Asn Gly Thr Ile 
            340                 345                 350         
Ser Trp Gly Ser Thr Ala Gly Phe Gly Asp Asp Val Asp Ile Phe Ala 
        355                 360                 365             
Glu Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu 
    370                 375                 380                 
Trp Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly 
385                 390                 395                 400 
Gln Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile 
                405                 410                 415     
Lys Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp 
            420                 425                 430         
Asp Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys 
        435                 440                 445             
Ala Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu 
    450                 455                 460                 
Thr Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val 
465                 470                 475                 480 
His Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu 
                485                 490                 495     
Pro Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe 
            500                 505                 510         
Asp Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn 
        515                 520                 525             
Trp Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala 
    530                 535                 540                 
Phe Leu Trp Gly Gly Ala Asp Arg Val Thr Glu Ile Asp Thr Ile Leu 
545                 550                 555                 560 
Asp Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg 
                565                 570                 575     
Gln Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu 
            580                 585                 590         
Lys Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu 
        595                 600                 605             
Val Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp 
    610                 615                 620                 
Gly Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu 
625                 630                 635                 640 
Thr Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe 
                645                 650                 655     
Gly Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro 
            660                 665                 670         
Thr Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala 
        675                 680                 685             
Leu Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly 
    690                 695                 700                 
Gly Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp 
705                 710                 715                 720 
Gln Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr 
                725                 730                 735     
Pro Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro 
            740                 745                 750         
Gln Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg 
        755                 760                 765             
Gly Thr Glu Asn Asp Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg 
    770                 775                 780                 
Pro Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile 
785                 790                 795                 800 
Ala Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met 
                805                 810                 815     
Tyr Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val 
            820                 825                 830         
Asp Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Arg         
        835                 840                 845             

 
<210> 3
<211> 2295
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 3
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgtaa                                                  2295

 
<210> 4
<211> 764
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 4
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala                 
        755                 760                                 

 
<210> 5
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 5
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 6
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 6
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 7
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 7
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 8
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 8
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 9
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 9
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtagtt 1080
gccgcggttc cagcgccgtt tggtaagtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 10
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 10
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 11
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 11
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagttt tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 12
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 12
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Phe Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 13
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 13
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagtgg tacagcatga gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 14
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 14
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 15
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 15
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtccgtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 16
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 16
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Pro Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 17
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 17
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tcgcgttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccag ctggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 18
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 18
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Ala Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Ala 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 19
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 19
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgcgcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt tccagttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggtcc cagcgccgtt tggtaagtgg tacagcgcca gtggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 20
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 20
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Phe Gln Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Trp Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 21
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 21
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgtggttc cagcgccgtt tggtaagatc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg aggcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagacg 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcgacagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 22
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 22
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Val Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Arg Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Asp Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 23
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 23
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tacgcggcgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcgacagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 24
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 24
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Asp Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 25
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 25
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggttc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtagggtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggcgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcatggaca ccatcaccat taa                              2313

 
<210> 26
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 26
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Phe Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Ala Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His Gly His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 27
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 27
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggttc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtagggtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccggcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcctcatca ccatcaccat taa                              2313

 
<210> 28
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 28
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Phe Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Gly Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala Pro His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 29
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 29
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgttg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggttc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtagggtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccggcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcatggaca ccatcaccat taa                              2313

 
<210> 30
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 30
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Phe Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Gly Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His Gly His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 31
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 31
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttcct gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtgtcttc tacagcgcct cgggctatga aaccacccag 1140
gacgggccaa ccggctcgct gaccatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcgcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgccgg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtagggtt ggatcagacc 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcatggaca ccatcaccat taa                              2313

 
<210> 32
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 32
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Val Phe Tyr Ser Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Thr Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Pro Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Thr 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His Gly His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 33
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 33
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttctt gtggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 34
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 34
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 35
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 35
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttctt gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatc tacgcgtttt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggttt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 36
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 36
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Phe Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 37
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 37
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttctt gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tactcgtttt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggtgt tgattcgggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 38
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 38
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ser Phe Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Val Asp Ser Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 39
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 39
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttctt gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tactcgtttt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggttt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcacgagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaaac agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 40
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 40
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ser Phe Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 41
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 41
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgttg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatctgt ggggcttctt gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggtaagatc tactcgtttt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacaagg tgatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcacgagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaaac agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 42
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 42
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Leu Trp Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Lys Ile Tyr Ser Phe Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 43
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 43
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcatc aaggtttcct ttggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatc tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gcgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaaggca ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 44
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 44
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Ala Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Gly Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 45
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 45
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcatc aaggtttcct ttggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatc tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaaggca ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 46
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 46
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Gly Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 47
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 47
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcatt cgggtttcct ttggggcggt 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gataaactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct taacgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatc tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcaccg gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaagttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcaagc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 48
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 48
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Ser Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Lys Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Asn Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Gly Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Lys Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 49
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 49
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacaggg ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaaggca ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 50
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 50
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Gly Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 51
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 51
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcgt gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccaa caggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggttt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 52
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 52
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Val Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Thr 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 53
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 53
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 54
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 54
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 55
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 55
agcaatatgt gggtgattgg caaaaaaaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgattctc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggtgg ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 56
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 56
Ser Asn Met Trp Val Ile Gly Lys Lys Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp Ser Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 57
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 57
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gcgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggtgg ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 58
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 58
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Ala Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 59
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 59
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcaagg agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gcgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggttt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 60
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 60
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Lys Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Ala Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 61
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 61
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gccggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggaaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaggtgg ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaaggca ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 62
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 62
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Glu Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gly Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Gly Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 63
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 63
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt cttggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aatgccgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcacttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acggcaaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacaggg tgatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg ccagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctggagaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 64
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 64
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Cys Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ala Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Glu Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 65
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 65
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt tttggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccggaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aatgccgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcacttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtctaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaatttt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg caagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctggacaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 66
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 66
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Phe Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Glu Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Cys Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Asn Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Asp Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 67
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 67
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt cttggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat taaaaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccggaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcggc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg catcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtctaaca acatgattgt cttctcaccg acgtcgggta accgcccggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt ggatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaact gcagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tatggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctggagaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 68
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 68
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile Lys 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Glu Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Gly Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met His Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Cys Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Glu Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 69
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 69
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt gcgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 70
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 70
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Ala Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 71
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 71
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taatccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtgcaaca acatgattgt cttctcaccg acgtcgggta accgcaaggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgtttcattt 1860
gataaagata ttcgccaaaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg caagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 72
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 72
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Cys Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Lys 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Ser Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 73
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 73
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 74
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 74
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 75
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 75
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcgatgt cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgcaaggt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccaaaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg caagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctatccgc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 76
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 76
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Met Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Lys 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Ala Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Pro Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 77
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 77
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aatgccgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgctgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgagcaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatggggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctggataac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 78
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 78
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Cys Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Trp Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Asp Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 79
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 79
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttcggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgacctcga tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg catcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtaaactt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt tgttaagttt 1860
gataaggata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaacagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc ggctgaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 80
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 80
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Phe Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met His Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Asn Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Asn Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Ala Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 81
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 81
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgctgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgagcaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacaggg tgatcagggt 1680
gaggttaaga tcgttcgcga tgaatggggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaaacttt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 82
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 82
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Gly Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Trp Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Thr Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 83
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 83
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgctgcctgg ctgaccagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctggataac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 84
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 84
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Asp Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 85
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 85
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgagcagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacattagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcagtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 86
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 86
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Ser 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 87
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 87
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt ttgggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac agccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgagcat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcaat cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgagcagca tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg gcccttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacagtt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt cgttaaattt 1860
gataaagata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg aaagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcactagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc gcgggaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 88
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 88
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Leu Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Ser Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Ser 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Ser 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met Ala Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Arg Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

 
<210> 89
<211> 2313
<212> DNA
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 89
agcaatatgt gggtgattgg caaaaacaaa gcccaggatg cgaaggccat tatggtcaat 60
gggccgcagt ttggttggta taacccggcg tatacctacg gtatcggcct gcacggcgcg 120
ggctatgacg tcaccggcaa tacgccgttt gcctatccgg gcctcctttt tggtcacaac 180
ggcaccattt catggggatc caccgccggt tgcggtgata gcgtcgatat ctttgccgaa 240
aaactttccg ccgagaagcc gggctattac cagcataacg gcgagtgggt gaagatgttg 300
agccgcgtag agactattgc ggtcaaagac ggccagccgg agacctttac cgtttggcgc 360
acgctgcacg gcaacgtcat tcacaccgat actgcgacgc agaccgccta tgccaaagcg 420
cgggcctggg atggcaaaga ggtggcgtcc ctgctggcgt ggacgcacca gatgaaggcc 480
aaaaactggc cggagtggac gcagcaggcg gccaaacagg cgctgaccat caactggtac 540
tacgccgatg tgaacggcaa tatcggctat gtgcataccg gcgcctatcc ggatcgccag 600
cccggccacg acccgcgttt gccggttccc ggcactggaa aatgggactg gaaagggttg 660
ctgtcgtttg atttgaatcc gaaagtgtat aacccgcagt cgggctatat cgccaactgg 720
aacaactcgc cgcaaaaaga ctacccggcc tctgatcacc aaggtttcct gtggggcagc 780
gcggatcgag cgactgagat cgacacgatc ctcgataagc aaccgcgctt caccgccgat 840
caggcgtggg atgtgatccg ccaaaccagc cgtcgggatc tcaacctgcg gttgttctta 900
ccggcgctga aggacgccac cgccaacctg gcggaaaacg atccgcgccg ccaactggtg 960
gatacactgg cgagctggga cggcgaaaac cttgtcaacg atgacggaaa aacctatcag 1020
caaccgggat cggcgattct tgcagcctgg ctgacctcga tgctcaagcg cacggtggtt 1080
gccgcggttc cagcgccgtt tggttggatt tacgcggcat cgggctatga aaccacccag 1140
gacgggccac tcggctcgct gaacatcagc gtgggggcga aaatcctcta cgaagctctg 1200
cagggtgata agtcgccaat cccgcaggcg gtcgatctgt ttggcgggaa accgcagcag 1260
gaagtaatac tggcggcgct ggacgacgct tggcagacgc tgtcaaaacg ctacggtaac 1320
gacgtcacca gctggaaaac ccctgccatg catcttacct tccgggccaa taacttcttc 1380
ggcgtgccgc aggcggcagc aaaagaggcg cgtcatcagg cggagtacca gaaccgcggt 1440
acgtccaaca acatgattgt cttctcaccg acgtcgggta accgccccgt tcttgcctgg 1500
gatgtggtgg cgccggggca aagcggtttt atcgcgcagg atggcaaagc cgataagcac 1560
tatgacgatc agctgaaaat gtacgagagc tttggccgta aatcgctgtg gttaacgcct 1620
caggacgttg acgagcacaa agagtctcag gaagtgctgc aggtacaatt agatcagggt 1680
gaggttaaga tcgttcgcga tgaatacggc atgccgcata tttacgccga tgatacctat 1740
cgactgtttt acggctatgg ctacgtggtg gcgcaggatc gcctgttcca gatggaaatg 1800
gcgcgccgca gtactcaggg gaccgtctcc gaggtgctgg gcaaagcatt tgttaagttt 1860
gataaggata ttcgccagaa ctactggccg gattctattc gcgcgcagat agcttccctc 1920
tccgctgagg ataaatccat tctgcagggc tatgccgatg gcatgaatgc gtggatcgat 1980
aaagtgaacg agagccccga taagctgtta ccccagcagt tctccacctt tggttttaaa 2040
cccaagcatt gggaaccgtt tgatgtggcg atgatttttg tcggcaccat ggcgaaccgt 2100
ttttctgaca gcaccagcga aattgataac ctggcgctgc tgacggcgct aaaagacaaa 2160
tacggcgcgc agcagggcat ggcggtcttt aaccagctga aatggctggt taatccttcc 2220
gcgccaacca ccattgcggc ggctgaaagc gcctattgcc tgaagtttga tctgcaaaac 2280
acgcaaacgg cgcaccatca ccatcaccat taa                              2313

 
<210> 90
<211> 770
<212> PRT
<213> Artificial Sequence
 
<220>
<223> Variant of Penicillin G Acylase From Kluyvera Citrophila
 
<400> 90
Ser Asn Met Trp Val Ile Gly Lys Asn Lys Ala Gln Asp Ala Lys Ala 
1               5                   10                  15      
Ile Met Val Asn Gly Pro Gln Phe Gly Trp Tyr Asn Pro Ala Tyr Thr 
            20                  25                  30          
Tyr Gly Ile Gly Leu His Gly Ala Gly Tyr Asp Val Thr Gly Asn Thr 
        35                  40                  45              
Pro Phe Ala Tyr Pro Gly Leu Leu Phe Gly His Asn Gly Thr Ile Ser 
    50                  55                  60                  
Trp Gly Ser Thr Ala Gly Cys Gly Asp Ser Val Asp Ile Phe Ala Glu 
65                  70                  75                  80  
Lys Leu Ser Ala Glu Lys Pro Gly Tyr Tyr Gln His Asn Gly Glu Trp 
                85                  90                  95      
Val Lys Met Leu Ser Arg Val Glu Thr Ile Ala Val Lys Asp Gly Gln 
            100                 105                 110         
Pro Glu Thr Phe Thr Val Trp Arg Thr Leu His Gly Asn Val Ile His 
        115                 120                 125             
Thr Asp Thr Ala Thr Gln Thr Ala Tyr Ala Lys Ala Arg Ala Trp Asp 
    130                 135                 140                 
Gly Lys Glu Val Ala Ser Leu Leu Ala Trp Thr His Gln Met Lys Ala 
145                 150                 155                 160 
Lys Asn Trp Pro Glu Trp Thr Gln Gln Ala Ala Lys Gln Ala Leu Thr 
                165                 170                 175     
Ile Asn Trp Tyr Tyr Ala Asp Val Asn Gly Asn Ile Gly Tyr Val His 
            180                 185                 190         
Thr Gly Ala Tyr Pro Asp Arg Gln Pro Gly His Asp Pro Arg Leu Pro 
        195                 200                 205             
Val Pro Gly Thr Gly Lys Trp Asp Trp Lys Gly Leu Leu Ser Phe Asp 
    210                 215                 220                 
Leu Asn Pro Lys Val Tyr Asn Pro Gln Ser Gly Tyr Ile Ala Asn Trp 
225                 230                 235                 240 
Asn Asn Ser Pro Gln Lys Asp Tyr Pro Ala Ser Asp His Gln Gly Phe 
                245                 250                 255     
Leu Trp Gly Ser Ala Asp Arg Ala Thr Glu Ile Asp Thr Ile Leu Asp 
            260                 265                 270         
Lys Gln Pro Arg Phe Thr Ala Asp Gln Ala Trp Asp Val Ile Arg Gln 
        275                 280                 285             
Thr Ser Arg Arg Asp Leu Asn Leu Arg Leu Phe Leu Pro Ala Leu Lys 
    290                 295                 300                 
Asp Ala Thr Ala Asn Leu Ala Glu Asn Asp Pro Arg Arg Gln Leu Val 
305                 310                 315                 320 
Asp Thr Leu Ala Ser Trp Asp Gly Glu Asn Leu Val Asn Asp Asp Gly 
                325                 330                 335     
Lys Thr Tyr Gln Gln Pro Gly Ser Ala Ile Leu Ala Ala Trp Leu Thr 
            340                 345                 350         
Ser Met Leu Lys Arg Thr Val Val Ala Ala Val Pro Ala Pro Phe Gly 
        355                 360                 365             
Trp Ile Tyr Ala Ala Ser Gly Tyr Glu Thr Thr Gln Asp Gly Pro Leu 
    370                 375                 380                 
Gly Ser Leu Asn Ile Ser Val Gly Ala Lys Ile Leu Tyr Glu Ala Leu 
385                 390                 395                 400 
Gln Gly Asp Lys Ser Pro Ile Pro Gln Ala Val Asp Leu Phe Gly Gly 
                405                 410                 415     
Lys Pro Gln Gln Glu Val Ile Leu Ala Ala Leu Asp Asp Ala Trp Gln 
            420                 425                 430         
Thr Leu Ser Lys Arg Tyr Gly Asn Asp Val Thr Ser Trp Lys Thr Pro 
        435                 440                 445             
Ala Met His Leu Thr Phe Arg Ala Asn Asn Phe Phe Gly Val Pro Gln 
    450                 455                 460                 
Ala Ala Ala Lys Glu Ala Arg His Gln Ala Glu Tyr Gln Asn Arg Gly 
465                 470                 475                 480 
Thr Ser Asn Asn Met Ile Val Phe Ser Pro Thr Ser Gly Asn Arg Pro 
                485                 490                 495     
Val Leu Ala Trp Asp Val Val Ala Pro Gly Gln Ser Gly Phe Ile Ala 
            500                 505                 510         
Gln Asp Gly Lys Ala Asp Lys His Tyr Asp Asp Gln Leu Lys Met Tyr 
        515                 520                 525             
Glu Ser Phe Gly Arg Lys Ser Leu Trp Leu Thr Pro Gln Asp Val Asp 
    530                 535                 540                 
Glu His Lys Glu Ser Gln Glu Val Leu Gln Val Gln Leu Asp Gln Gly 
545                 550                 555                 560 
Glu Val Lys Ile Val Arg Asp Glu Tyr Gly Met Pro His Ile Tyr Ala 
                565                 570                 575     
Asp Asp Thr Tyr Arg Leu Phe Tyr Gly Tyr Gly Tyr Val Val Ala Gln 
            580                 585                 590         
Asp Arg Leu Phe Gln Met Glu Met Ala Arg Arg Ser Thr Gln Gly Thr 
        595                 600                 605             
Val Ser Glu Val Leu Gly Lys Ala Phe Val Lys Phe Asp Lys Asp Ile 
    610                 615                 620                 
Arg Gln Asn Tyr Trp Pro Asp Ser Ile Arg Ala Gln Ile Ala Ser Leu 
625                 630                 635                 640 
Ser Ala Glu Asp Lys Ser Ile Leu Gln Gly Tyr Ala Asp Gly Met Asn 
                645                 650                 655     
Ala Trp Ile Asp Lys Val Asn Glu Ser Pro Asp Lys Leu Leu Pro Gln 
            660                 665                 670         
Gln Phe Ser Thr Phe Gly Phe Lys Pro Lys His Trp Glu Pro Phe Asp 
        675                 680                 685             
Val Ala Met Ile Phe Val Gly Thr Met Ala Asn Arg Phe Ser Asp Ser 
    690                 695                 700                 
Thr Ser Glu Ile Asp Asn Leu Ala Leu Leu Thr Ala Leu Lys Asp Lys 
705                 710                 715                 720 
Tyr Gly Ala Gln Gln Gly Met Ala Val Phe Asn Gln Leu Lys Trp Leu 
                725                 730                 735     
Val Asn Pro Ser Ala Pro Thr Thr Ile Ala Ala Ala Glu Ser Ala Tyr 
            740                 745                 750         
Cys Leu Lys Phe Asp Leu Gln Asn Thr Gln Thr Ala His His His His 
        755                 760                 765             
His His                                                         
    770                                                         

