               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> Proline specific endoprotease

<130> 30253-WO-PCT

<160> 8

<170> BiSSAP 1.2

<210> 1
<211> 1400
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1400
<223> /organism="Artificial Sequence"
      /note="Codon optimized nucleic acid encoding proline-specific
      endoprotease BC2G076 Aspergillus_aculeatus, for expression in A.
      niger"
      /mol_type="unassigned DNA"

<400> 1
atggtcaagt ccattcttgc ctccgtcttc ttcgctgcca ctgctcttgc tgctcgtccc      60

cgtctcgtcc ccaagcccgt ctcccgcccc gccagcagca agtctgctgc caccaccggt     120

gaagcctact tcgagcagct gatcgaccac tccgacccct ccaagggtac tttctcccag     180

cgctactggt actctgctca gtactggggt ggtcctggca gccctgtcgt cctcttcact     240

cctggtgaag tgtctgccga tggctaccag ggctacctca ccaacgccac cctgaccggt     300

gtctacgctc agcagctcca gggtgccgtt gtcctcgtcg agcaccgcta ctggggcggc     360

agctctccct acaccaactt gactgctgag actctccagt acttgacttt ggagcagagc     420

gtgcttgacc tgacctactt cgctgagaac gtcaagctcg gtttcgacaa ctcgacctcc     480

tccaacgctc ctcacgtccc ctgggtgctg gtcggtggaa gctactctgg tgctctgacc     540

gcctggaccg aacacctggc tcctggcacc ttctgggcct accacgccac ctcggctccc     600

gtggagagca tctacgactt ctggcagtac ttccgtccca tccaggatgg catggccaag     660

aactgctcca aggatgtctc gctagttgct gaacacgttg acaagatcgg caagaccggc     720

accaaggccc agcagaccga gctgaagaag ctcttcggtc tggaagccct tgaacacttc     780

gatgacttcg ctgctgtcct tcccattggt ccctacctct ggcaggacaa caccttcgcc     840

actggatact ccgacttctt cgccttctgt gatgccgttg agaacgttga ggctggtgct     900

gctgtcaccc ccggtgctga gggtgttggt cttgagaagg ccctcaccgg atacgccaac     960

tggttcaaga acgagatctt ccccggatac tgcgcttcct acggctactg gtcagatgag    1020

tactctgttg cctgctacga cacctacaac accacttctc ccctcttcac cgacacctcc    1080

gtcgacaacg ccgttgaccg ccagtggcag tggttcctgt gcaacgagcc cttcttctgg    1140

tggcaggatg gtgctccctc ctccgagact accattgtgc ctcgtctcgt ctctgccgac    1200

tactggcagc gccagtgcgc tctctacttc cccgaggtca acggatacac ctacggctct    1260

gccaagggca agtccgccaa cactttcaac gcctggactg atggctggtt catgaatggc    1320

aacagcaccc gtctgatctg gaccaacggc cagtacgacc cttggcgtga tgccaccgtc    1380

tcctccacct tccgccccgg                                                1400


<210> 2
<211> 521
<212> PRT
<213> Artificial Sequence

<220> 
<223> Proline-specific endoprotease- BC2G076_Aspergillus_aculeatus

<400> 2
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser 
            20                  25                  30          
Ser Lys Ser Ala Ala Thr Thr Gly Glu Ala Tyr Phe Glu Gln Leu Ile 
        35                  40                  45              
Asp His Ser Asp Pro Ser Lys Gly Thr Phe Ser Gln Arg Tyr Trp Tyr 
    50                  55                  60                  
Ser Ala Gln Tyr Trp Gly Gly Pro Gly Ser Pro Val Val Leu Phe Thr 
65                  70                  75                  80  
Pro Gly Glu Val Ser Ala Asp Gly Tyr Gln Gly Tyr Leu Thr Asn Ala 
                85                  90                  95      
Thr Leu Thr Gly Val Tyr Ala Gln Gln Leu Gln Gly Ala Val Val Leu 
            100                 105                 110         
Val Glu His Arg Tyr Trp Gly Gly Ser Ser Pro Tyr Thr Asn Leu Thr 
        115                 120                 125             
Ala Glu Thr Leu Gln Tyr Leu Thr Leu Glu Gln Ser Val Leu Asp Leu 
    130                 135                 140                 
Thr Tyr Phe Ala Glu Asn Val Lys Leu Gly Phe Asp Asn Ser Thr Ser 
145                 150                 155                 160 
Ser Asn Ala Pro His Val Pro Trp Val Leu Val Gly Gly Ser Tyr Ser 
                165                 170                 175     
Gly Ala Leu Thr Ala Trp Thr Glu His Leu Ala Pro Gly Thr Phe Trp 
            180                 185                 190         
Ala Tyr His Ala Thr Ser Ala Pro Val Glu Ser Ile Tyr Asp Phe Trp 
        195                 200                 205             
Gln Tyr Phe Arg Pro Ile Gln Asp Gly Met Ala Lys Asn Cys Ser Lys 
    210                 215                 220                 
Asp Val Ser Leu Val Ala Glu His Val Asp Lys Ile Gly Lys Thr Gly 
225                 230                 235                 240 
Thr Lys Ala Gln Gln Thr Glu Leu Lys Lys Leu Phe Gly Leu Glu Ala 
                245                 250                 255     
Leu Glu His Phe Asp Asp Phe Ala Ala Val Leu Pro Ile Gly Pro Tyr 
            260                 265                 270         
Leu Trp Gln Asp Asn Thr Phe Ala Thr Gly Tyr Ser Asp Phe Phe Ala 
        275                 280                 285             
Phe Cys Asp Ala Val Glu Asn Val Glu Ala Gly Ala Ala Val Thr Pro 
    290                 295                 300                 
Gly Ala Glu Gly Val Gly Leu Glu Lys Ala Leu Thr Gly Tyr Ala Asn 
305                 310                 315                 320 
Trp Phe Lys Asn Glu Ile Phe Pro Gly Tyr Cys Ala Ser Tyr Gly Tyr 
                325                 330                 335     
Trp Ser Asp Glu Tyr Ser Val Ala Cys Tyr Asp Thr Tyr Asn Thr Thr 
            340                 345                 350         
Ser Pro Leu Phe Thr Asp Thr Ser Val Asp Asn Ala Val Asp Arg Gln 
        355                 360                 365             
Trp Gln Trp Phe Leu Cys Asn Glu Pro Phe Phe Trp Trp Gln Asp Gly 
    370                 375                 380                 
Ala Pro Ser Ser Glu Thr Thr Ile Val Pro Arg Leu Val Ser Ala Asp 
385                 390                 395                 400 
Tyr Trp Gln Arg Gln Cys Ala Leu Tyr Phe Pro Glu Val Asn Gly Tyr 
                405                 410                 415     
Thr Tyr Gly Ser Ala Lys Gly Lys Ser Ala Asn Thr Phe Asn Ala Trp 
            420                 425                 430         
Thr Asp Gly Trp Phe Met Asn Gly Asn Ser Thr Arg Leu Ile Trp Thr 
        435                 440                 445             
Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ala Thr Val Ser Ser Thr Phe 
    450                 455                 460                 
Arg Pro Gly Gly Pro Leu Ala Ser Thr Pro Ser Glu Pro Val Gln Ile 
465                 470                 475                 480 
Ile Pro Gly Gly Phe His Cys Ser Asp Leu Tyr Ile Ser Asp Ser Val 
                485                 490                 495     
Val Asn Ala Gly Val Lys Lys Val Val Asp Asn Glu Val Ala Gln Ile 
            500                 505                 510         
Lys Ala Trp Val Ala Glu Phe Tyr Ala 
        515                 520     

<210> 3
<211> 17
<212> PRT
<213> Artificial Sequence

<220> 
<223> Signal sequence of A. niger pectinemethylesterase PMeA ss

<400> 3
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala 
    

<210> 4
<211> 19
<212> PRT
<213> Artificial Sequence

<220> 
<223> Pro-sequence A.niger proline-specific endoprotease

<400> 4
Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser Ser 
1               5                   10                  15      
Lys Ser Ala 
            

<210> 5
<211> 521
<212> PRT
<213> Artificial Sequence

<220> 
<223> Proline-specific endoprotease from Aspergillus niger comprising a
      PMeA signal sequence from A. niger

<400> 5
Met Val Lys Ser Ile Leu Ala Ser Val Phe Phe Ala Ala Thr Ala Leu 
1               5                   10                  15      
Ala Ala Arg Pro Arg Leu Val Pro Lys Pro Val Ser Arg Pro Ala Ser 
            20                  25                  30          
Ser Lys Ser Ala Ala Thr Thr Gly Glu Ala Tyr Phe Glu Gln Leu Leu 
        35                  40                  45              
Asp His His Asn Pro Glu Lys Gly Thr Phe Ser Gln Arg Tyr Trp Trp 
    50                  55                  60                  
Ser Thr Glu Tyr Trp Gly Gly Pro Gly Ser Pro Val Val Leu Phe Thr 
65                  70                  75                  80  
Pro Gly Glu Val Ser Ala Asp Gly Tyr Glu Gly Tyr Leu Thr Asn Glu 
                85                  90                  95      
Thr Leu Thr Gly Val Tyr Ala Gln Glu Ile Gln Gly Ala Val Ile Leu 
            100                 105                 110         
Ile Glu His Arg Tyr Trp Gly Asp Ser Ser Pro Tyr Glu Val Leu Asn 
        115                 120                 125             
Ala Glu Thr Leu Gln Tyr Leu Thr Leu Asp Gln Ala Ile Leu Asp Met 
    130                 135                 140                 
Thr Tyr Phe Ala Glu Thr Val Lys Leu Gln Phe Asp Asn Ser Thr Arg 
145                 150                 155                 160 
Ser Asn Ala Gln Asn Ala Pro Trp Val Met Val Gly Gly Ser Tyr Ser 
                165                 170                 175     
Gly Ala Leu Thr Ala Trp Thr Glu Ser Val Ala Pro Gly Thr Phe Trp 
            180                 185                 190         
Ala Tyr His Ala Thr Ser Ala Pro Val Glu Ala Ile Tyr Asp Tyr Trp 
        195                 200                 205             
Gln Tyr Phe Tyr Pro Ile Gln Gln Gly Met Ala Gln Asn Cys Ser Lys 
    210                 215                 220                 
Asp Val Ser Leu Val Ala Glu Tyr Val Asp Lys Ile Gly Lys Asn Gly 
225                 230                 235                 240 
Thr Ala Lys Glu Gln Gln Ala Leu Lys Glu Leu Phe Gly Leu Gly Ala 
                245                 250                 255     
Val Glu His Phe Asp Asp Phe Ala Ala Val Leu Pro Asn Gly Pro Tyr 
            260                 265                 270         
Leu Trp Gln Asp Asn Asp Phe Ala Thr Gly Tyr Ser Ser Phe Phe Gln 
        275                 280                 285             
Phe Cys Asp Ala Val Glu Gly Val Glu Ala Gly Ala Ala Val Thr Pro 
    290                 295                 300                 
Gly Pro Glu Gly Val Gly Leu Glu Lys Ala Leu Ala Asn Tyr Ala Asn 
305                 310                 315                 320 
Trp Phe Asn Ser Thr Ile Leu Pro Asp Tyr Cys Ala Ser Tyr Gly Tyr 
                325                 330                 335     
Trp Thr Asp Glu Trp Ser Val Ala Cys Phe Asp Ser Tyr Asn Ala Ser 
            340                 345                 350         
Ser Pro Ile Tyr Thr Asp Thr Ser Val Gly Asn Ala Val Asp Arg Gln 
        355                 360                 365             
Trp Glu Trp Phe Leu Cys Asn Glu Pro Phe Phe Tyr Trp Gln Asp Gly 
    370                 375                 380                 
Ala Pro Glu Gly Thr Ser Thr Ile Val Pro Arg Leu Val Ser Ala Ser 
385                 390                 395                 400 
Tyr Trp Gln Arg Gln Cys Pro Leu Tyr Phe Pro Glu Thr Asn Gly Tyr 
                405                 410                 415     
Thr Tyr Gly Ser Ala Lys Gly Lys Asn Ala Ala Thr Val Asn Ser Trp 
            420                 425                 430         
Thr Gly Gly Trp Asp Met Thr Arg Asn Thr Thr Arg Leu Ile Trp Thr 
        435                 440                 445             
Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ser Gly Val Ser Ser Thr Phe 
    450                 455                 460                 
Arg Pro Gly Gly Pro Leu Ala Ser Thr Ala Asn Glu Pro Val Gln Ile 
465                 470                 475                 480 
Ile Pro Gly Gly Phe His Cys Ser Asp Leu Tyr Met Ala Asp Tyr Tyr 
                485                 490                 495     
Ala Asn Glu Gly Val Lys Lys Val Val Asp Asn Glu Val Lys Gln Ile 
            500                 505                 510         
Lys Glu Trp Val Glu Glu Tyr Tyr Ala 
        515                 520     

<210> 6
<211> 1566
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1566
<223> /organism="Artificial Sequence"
      /note="Proline-specific endoprotease from Aspergillus niger
      comprising a PMeA signal sequence from A. niger"
      /mol_type="unassigned DNA"

<400> 6
atggtcaagt ccatcctggc ctccgtcttc ttcgctgcca ctgctcttgc tgcaaggcct      60

cgtctcgttc ccaagcccgt ttctcgtccc gccagctcca agtccgctgc tactactggt     120

gaggcctact ttgaacagct gttggaccac cacaaccctg agaagggtac tttctcgcaa     180

agatactggt ggagcaccga gtactggggt ggtcccggat cccccgttgt cctgttcact     240

cccggtgagg tcagcgctga tggctacgag ggttatctga ccaacgagac tctcaccggt     300

gtctacgccc aggagattca gggtgctgtc atcctgatcg aacaccgata ctggggtgac     360

tcgtctccct acgaggtgct gaacgccgag actctccagt acttgaccct cgaccaggct     420

atccttgata tgacctactt cgccgaaacc gtcaagctcc agtttgacaa ctccacccgc     480

tccaacgctc agaacgctcc ttgggttatg gtcggcggca gctacagcgg tgctctgact     540

gcttggaccg agtccgttgc tcccggcacc ttctgggctt accacgccac ctctgctcct     600

gttgaggcca tctacgacta ctggcaatac ttctacccca ttcagcaggg tatggctcag     660

aactgctcca aagatgtctc tcttgtagca gaatacgtcg acaagatcgg caagaacggc     720

actgccaagg agcaacaggc tctgaaggag cttttcggcc taggagcagt ggagcacttc     780

gacgacttcg ccgctgttct gcccaacggt ccttacctct ggcaagacaa cgactttgcc     840

accggttact cttctttctt ccagttctgt gatgccgtcg agggtgtcga ggctggtgct     900

gccgtcaccc ccggtcctga aggtgttggt ctggaaaagg cccttgctaa ctacgcgaac     960

tggttcaact ctaccatcct ccccgattac tgcgccagct acggctactg gactgacgag    1020

tggtccgtcg cctgcttcga ctcctacaac gcctcctctc ctatatacac cgacaccagc    1080

gttggtaacg ccgtcgaccg tcagtgggag tggttcctct gcaatgagcc cttcttttac    1140

tggcaggacg gtgcccccga gggtacttca acgatagtac cccgcttagt gtccgcctcc    1200

tactggcagc gtcaatgtcc gttgtacttc cccgagacta acggttacac ctacggctcc    1260

gccaagggaa agaacgccgc caccgtcaac agctggaccg gtggctggga catgacccgt    1320

aacaccaccc gtctgatctg gacgaacggc caatacgacc cctggcgtga ctccggtgtc    1380

tcttccacct tccgccccgg tggtcccctc gcttcgaccg ccaacgagcc cgtccagata    1440

atacccggtg gtttccattg ctccgacctc tacatggcag actactacgc caacgagggc    1500

gtcaagaagg ttgtcgacaa cgaagtcaaa caaatcaagg agtgggttga ggaatactac    1560

gcgtaa                                                               1566


<210> 7
<211> 527
<212> PRT
<213> Aspergillus aculeatus

<220> 
<223> PEP BC2G112

<400> 7
Met Arg Phe Cys Ser Pro Val Ser Leu Val Ala Val Thr Ala Leu Trp 
1               5                   10                  15      
Ala Ser Leu Ala Ala Ala Val Arg Pro Pro Arg Leu Ala Pro Arg Pro 
            20                  25                  30          
Ile Ala Ala Thr Ser Gln Ala Ser Glu Ser Ser Val Val Glu Ser Thr 
        35                  40                  45              
Phe Glu Gln Leu Ile Asp His Ser Asp Pro Ser Lys Gly Thr Phe Ser 
    50                  55                  60                  
Gln Arg Tyr Trp Tyr Ser Ala Gln Tyr Trp Gly Gly Pro Gly Ser Pro 
65                  70                  75                  80  
Val Val Leu Phe Thr Pro Gly Glu Val Ser Ala Asp Gly Tyr Gln Gly 
                85                  90                  95      
Tyr Leu Thr Asn Ala Thr Leu Thr Gly Val Tyr Ala Gln Gln Leu Gln 
            100                 105                 110         
Gly Ala Val Val Leu Val Glu His Arg Tyr Trp Gly Gly Ser Ser Pro 
        115                 120                 125             
Tyr Thr Asn Leu Thr Ala Glu Thr Leu Gln Tyr Leu Thr Leu Glu Gln 
    130                 135                 140                 
Ser Val Leu Asp Leu Thr Tyr Phe Ala Glu Asn Val Lys Leu Gly Phe 
145                 150                 155                 160 
Ala Arg Asn Ser Thr Ser Ser Asn Ala Pro His Val Pro Trp Val Leu 
                165                 170                 175     
Val Gly Gly Ser Tyr Ser Gly Ala Leu Thr Ala Trp Thr Glu His Leu 
            180                 185                 190         
Ala Pro Gly Thr Phe Trp Ala Tyr His Ala Thr Ser Ala Pro Val Glu 
        195                 200                 205             
Ser Ile Tyr Asp Phe Trp Gln Tyr Phe Arg Pro Ile Gln Asp Gly Met 
    210                 215                 220                 
Ala Lys Asn Cys Ser Lys Asp Val Ser Leu Val Ala Glu His Val Asp 
225                 230                 235                 240 
Lys Ile Gly Lys Thr Gly Thr Lys Ala Gln Gln Thr Glu Leu Lys Lys 
                245                 250                 255     
Leu Phe Gly Leu Glu Ala Leu Glu His Phe Asp Asp Phe Ala Ala Val 
            260                 265                 270         
Leu Pro Ile Gly Pro Tyr Leu Trp Gln Asp Asn Thr Phe Ala Thr Gly 
        275                 280                 285             
Tyr Ser Asp Phe Phe Ala Phe Cys Asp Ala Val Glu Asn Val Glu Ala 
    290                 295                 300                 
Gly Ala Ala Val Thr Pro Gly Ala Glu Gly Val Gly Leu Glu Lys Ala 
305                 310                 315                 320 
Leu Thr Gly Tyr Ala Asn Trp Phe Lys Asn Glu Ile Phe Pro Gly Tyr 
                325                 330                 335     
Cys Ala Ser Tyr Gly Tyr Trp Ser Asp Glu Tyr Ser Val Ala Cys Tyr 
            340                 345                 350         
Asp Thr Tyr Asn Thr Thr Ser Pro Leu Phe Thr Asp Thr Ser Val Asp 
        355                 360                 365             
Asn Ala Val Asp Arg Gln Trp Gln Trp Phe Leu Cys Asn Glu Pro Phe 
    370                 375                 380                 
Phe Trp Trp Gln Asp Gly Ala Pro Ser Ser Glu Thr Thr Ile Val Pro 
385                 390                 395                 400 
Arg Leu Val Ser Ala Asp Tyr Trp Gln Arg Gln Cys Ala Leu Tyr Phe 
                405                 410                 415     
Pro Glu Val Asn Gly Tyr Thr Tyr Gly Ser Ala Lys Gly Lys Ser Ala 
            420                 425                 430         
Asn Thr Phe Asn Ala Trp Thr Asp Gly Trp Phe Met Asn Gly Asn Ser 
        435                 440                 445             
Thr Arg Leu Ile Trp Thr Asn Gly Gln Tyr Asp Pro Trp Arg Asp Ala 
    450                 455                 460                 
Thr Val Ser Ser Thr Phe Arg Pro Gly Gly Pro Leu Ala Ser Thr Pro 
465                 470                 475                 480 
Ser Glu Pro Val Gln Ile Ile Pro Gly Gly Phe His Cys Ser Asp Leu 
                485                 490                 495     
Tyr Ile Ser Asp Ser Val Val Asn Ala Gly Val Lys Lys Val Val Asp 
            500                 505                 510         
Asn Glu Val Ala Gln Ile Lys Ala Trp Val Ala Glu Phe Tyr Ala 
        515                 520                 525         

<210> 8
<211> 1581
<212> DNA
<213> Artificial Sequence

<220> 
<221> source
<222> 1..1581
<223> /organism="Artificial Sequence"
      /note="BC2G112 PEP cpo for expression in A. niger"
      /mol_type="unassigned DNA"

<400> 8
atgcgcttct gctctcctgt ctccctggtg gccgtcactg ctctctgggc gtcgctagct      60

gctgctgtcc gccctcctcg tctggctcct cgtcccattg ctgccacctc ccaggcctcc     120

gagagctctg ttgttgagtc caccttcgag cagctcattg accactccga cccctccaag     180

ggtaccttct cccagcgcta ctggtactct gctcagtact ggggcggtcc tggaagcccc     240

gtcgtcctct tcacccccgg tgaagtctct gccgatggat accagggcta cctcaccaac     300

gccaccctca ctggtgtcta cgcccagcag ctccagggtg ctgttgtcct tgttgaacac     360

cgttactggg gtggaagctc tccctacacc aacttgactg cggaaaccct gcagtacctg     420

accctcgagc agtccgtcct tgacctgacc tacttcgctg agaacgtcaa gctcggtttc     480

gctcgcaact cgaccagcag caacgctcct cacgttccct gggtgcttgt tggtggcagc     540

tactctggtg ctttgactgc ctggactgag cacctggctc ccggcacctt ctgggcctac     600

cacgccacct ccgctcctgt ggaatcgatc tacgacttct ggcagtactt ccgtcccatc     660

caggatggaa tggccaagaa ctgctccaag gatgtctctc ttgttgctga gcacgttgac     720

aagatcggca agaccggtac caaggcccag cagaccgagc tcaagaagct gttcggtctg     780

gaggctcttg agcacttcga tgacttcgct gccgtcctcc ccattggtcc ctacctctgg     840

caggacaaca ccttcgccac tggctactcc gacttcttcg ctttctgcga tgccgtcgag     900

aacgtcgagg ctggtgctgc tgtgaccccc ggcgctgagg gtgttggtct ggagaaggcc     960

ttgactggct acgccaactg gttcaagaac gagatcttcc ccggatactg cgccagctac    1020

ggctactggt cggatgagta ctctgttgcc tgctacgaca cctacaacac cacctcgcct    1080

ctcttcaccg acacctcggt ggacaacgcc gttgaccgcc agtggcagtg gttcctttgc    1140

aacgagccct tcttctggtg gcaggatggt gctccctcct ccgagactac catcgtcccc    1200

cgtctggtca gcgccgacta ctggcagcgc cagtgcgctc tctacttccc cgaagtcaac    1260

ggctacacct acggctctgc caagggcaag tccgccaaca ctttcaacgc ctggaccgat    1320

ggctggttca tgaatggcaa ctccacccgt ctgatctgga ccaacggcca gtacgacccc    1380

tggcgcgatg ccactgtctc cagcactttc cgtcctggtg gtcctcttgc cagcactccc    1440

tccgagcccg tgcagatcat ccccggtggt ttccactgct ccgacctcta catctccgac    1500

tccgtcgtca acgccggtgt caagaaggtc gtcgacaacg aggttgccca gatcaaggcc    1560

tgggtggcgg aattctatgc c                                              1581


