                         SEQUENCE LISTING

<110>  CHINA SINOPHARM INTERNATIONAL CORPORATION
       Nanjing Redwood Fine Chemical Co.,LTD
 
<120>  Method for biocatalytic synthesis of Sitagliptin and intermediate thereof

<130>  IEC180069PCT

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  343
<212>  PRT
<213>  artificial

<220>
<223>  the amino acid sequence of Arthrobacter-derived transaminase mutant 1

<400>  1

Ser Val Leu His Arg Gly Gln Gln Arg Arg Arg Phe His Ile Gln Phe 
1               5                   10                  15      


Pro Val Thr Thr Asp Asn Ala Leu Gly Asn Arg Thr Arg His Thr Val 
            20                  25                  30          


Arg Asn Gly Ile Thr Ile Asp Arg Asn Glu Arg Pro Asp Ala Thr Ala 
        35                  40                  45              


Gly Gly Ala Thr Gln Asn Phe Val Ser Ile Val Gln Phe Cys Arg Arg 
    50                  55                  60                  


Asp Ile Gly Gln Asn Arg Phe Val Ala Gln Arg Phe Arg Asp Phe Gln 
65                  70                  75                  80  


Asn Gly Leu Ala Arg Asn Thr Arg Gln Ser Cys Thr Thr Arg Ala Thr 
                85                  90                  95      


Asn His Thr Ile Phe Asp Asp Asn His Ile Lys Ala Arg Thr Phe Ser 
            100                 105                 110         


Gln Gln Val Val Thr Ile Gln Gln Gln Arg Gln Phe Glu Thr Ala Ile 
        115                 120                 125             


Met Gly Phe Leu Asp Cys Thr Asn Gln Val Ala Pro Leu Lys Val Leu 
    130                 135                 140                 


His Leu Arg Ile Asn Gly Thr Thr Arg Gly Ala Thr Asp Ala Leu Cys 
145                 150                 155                 160 


Asn His Gln Val His Thr Val Ala Asp Thr Ile Glu Arg Asn Asn Pro 
                165                 170                 175     


Leu Val Arg Ala Arg Thr His Ile His Leu Arg Ala Met Phe Gly Asp 
            180                 185                 190         


Ile Thr Phe Lys Arg Arg Arg Ala Ile Ala Ala Gly Asn Arg His Gly 
        195                 200                 205             


Asp His Gly Phe Thr Gln Phe Gly Leu Gly His Gln Phe Gln Arg Asp 
    210                 215                 220                 


Phe Phe Asp Phe Ile Leu Arg Gln Arg Arg Asp Gln Ala Asn Arg Phe 
225                 230                 235                 240 


Cys Ile Ala Glu Gln Ala Phe Asn Val Val Ala Gln Thr Glu Ser Ile 
                245                 250                 255     


Thr Val Pro Asn Met Lys Arg Gly Val Gly Cys Val Arg Arg Ile Glu 
            260                 265                 270         


Thr Leu Ile Lys Asp Gly Asn Thr Gly Phe Thr Arg Arg His Lys Arg 
        275                 280                 285             


Thr Leu Asn Pro Cys Cys Thr Ala Ser Gln Arg Val Cys Arg Val Gln 
    290                 295                 300                 


Phe Val Val Ala Val Gly Asn Val Val Gln Ala Arg Ile Val Gly Val 
305                 310                 315                 320 


Asn Asn Phe Arg Arg Val Cys Gly Glu Cys His Arg Ile Leu Ala Val 
                325                 330                 335     


Val Met Met Val Met Ala Ala 
            340             


<210>  2
<211>  972
<212>  DNA
<213>  artificial

<220>
<223>  the gene sequence of Arthrobacter-derived transaminase mutant 1

<400>  2
atggcctcta tggacaaagt cttttcggga tattatgcgc gccagaagct gcttgaacgg       60

agcgacaatc ctttctctaa gggcattgct tatgtggaag gaaagctcgt ctttcctagt      120

gatgctagaa taccgctact cgacgaaggt ttcatgcaca gtgacctaac ctatgatgtt      180

atatcggttt gggatggtcg cttctttcga ttggacgatc atttgcaacg gattttggaa      240

agctgcgata agatgcggct caagttccca cttgcactga gcaccgtgga aaatattctg      300

gctgagatgg tcgccaagag tggtatccgg gatgcgtttg tggaagttat tgtgacacgt      360

ggtctgacag gtgtacgtgg ttcgaagcct gaggatctgt ataataacaa catatacctg      420

cttgttcttc catacatttg ggttatggcg cctgagaacc agctccatgg tggcgaggct      480

atcattacaa ggacagtgcg acgaacaccc ccaggtgcat ttgatcctac tatcaaaaat      540

ctacagtggg gtgatttaac aaagggactt tttgaggcaa tggaccgtgg cgccacatac      600

ccatttctca ctgatggaga caccaacctt actgaaggat ctggtttcaa cattgttttg      660

gtgaagaacg gtattatcta tacccctgat cgaggtgtct tgcgagggat cacacgtaaa      720

agtgtgattg acgttgcccg agccaacagc atcgacatcc gccttgaggt cgtaccagtg      780

gagcaggctt atcactctga tgagatcttc atgtgcacaa ctgccggcgg cattatgcct      840

ataacattgc ttgatggtca acctgttaat gacggccagg ttggcccaat cacaaagaag      900

atatgggatg gctattggga gatgcactac aatccggcgt atagttttcc tgttgactat      960

ggcagtggct aa                                                          972


<210>  3
<211>  322
<212>  PRT
<213>  artificial

<220>
<223>  the amino acid sequence of Arthrobacter-derived transaminase mutant 2

<400>  3

Ala Ala Ala Ile Thr Ile Ile Thr Thr Ala Arg Ile Arg Tyr Cys Thr 
1               5                   10                  15      


Gly Val Ser Arg Glu Glu Asp Ser Thr Phe Ser Ser Gln Gly Arg Arg 
            20                  25                  30          


Met Ile Asp Trp Val Thr Gly Pro Gly Thr Pro Ser Glu Ile Gly Leu 
        35                  40                  45              


Pro Ser Thr Glu Thr Asn Gly Gln Thr Pro Pro Pro Val Val Gln Pro 
    50                  55                  60                  


Arg Thr Ser Ser Ala Ser Ser Ser Ser Ala Arg Val Met Ser Ala Arg 
65                  70                  75                  80  


Ile Ala Ser Gly Pro Arg Asp Ser Ala Ile Ser Arg Thr Val Leu Arg 
                85                  90                  95      


Val Ile Pro Gly Arg Ala Ala Ala Ser Arg Pro Ser Pro Ser Ser Ser 
            100                 105                 110         


Ser Gly Ala Ser Lys Pro Arg Ser Trp Val Ser Gly Thr Ala Arg Ile 
        115                 120                 125             


Arg Ser Pro His Trp Lys Phe Leu Thr Cys Gly Ser Ile Glu Glu Arg 
    130                 135                 140                 


Gly Val Arg Arg Thr Asp Gly Ala Thr Ile Ala Gly Thr Pro Ser Arg 
145                 150                 155                 160 


Met Arg Ser Asn Gly Thr Ile His Trp Tyr Gly Thr Ala Tyr Met Gly 
                165                 170                 175     


Thr Cys Gly Arg Cys Leu Val Met Ser Arg Ser Pro Gly Val Glu Glu 
            180                 185                 190         


Gly Pro Arg Val Ile Glu Thr Glu Thr Thr Ala Ser Arg Ser Ser Val 
        195                 200                 205             


Leu Ala Thr Ser Ser Arg Ala Ile Ser Leu Thr Ser Ser Trp Val Ser 
    210                 215                 220                 


Gly Gly Met Ile Arg Ile Asp Ser Ala Leu Glu Asn Arg Arg Ser Met 
225                 230                 235                 240 


Trp Ser Ser Arg Arg Lys Ala Leu Pro Phe Gln Thr Trp Asn Pro Val 
                245                 250                 255     


Gly Val Thr Ser Glu Cys Arg Gly Pro Trp Ser Lys Ile Glu Ile Arg 
            260                 265                 270         


Ala Ser Asp Gly Gly Thr Lys Ala Pro Ser Ile Gln Ala Ala Pro Pro 
        275                 280                 285             


Ala Ser Gly Leu Ala Gly Ser Ser Ser Gly Ser Glu Gly Val Ile Gly 
    290                 295                 300                 


Ser Arg Pro Val Ser Trp Val Gly Thr Ile Ser Glu Val Ser Ala Glu 
305                 310                 315                 320 


Lys Ala 
        


<210>  4
<211>  968
<212>  DNA
<213>  artificial

<220>
<223>  the gene sequence of Arthrobacter-derived transaminase mutant 2

<400>  4
gcagcagcca tcaccatcat caccacagcc aggatccggt actgtaccgg ggtcagcaga       60

gaagaagatt caacgttcag ttcccagtaa cgacggatga tagactgggt aaccggaccc      120

ggaacaccgt cagagatcgg gttaccgtca acagaaacga acggccaaac accaccaccg      180

gtagtgcaac ccagaacttc gtcagcgtcc agcagttcag ccagggtgat gtcagccagg      240

atagcttcgt gacccagaga ttcagcgatt tccagaacgg ttttacgggt gatacccggc      300

agagcagcag ccagcagacc gtcaccgtcc agcagcagcg gagcttcgaa accacggtcg      360

tgggtttcct gaactgcacg gatcaggtca ccccactgga agtttttaac ctgcgggtcg      420

atagaagaac gcggagtacg acgaacagac tgagcaacca tagcgtgaac accgtcacgg      480

atgcggtcaa acggtacgat ccactggtac ggaacagcgt acatgtaaac ctgcggacga      540

tgtttggtga tgtcacgttc acctggggta gaagagtaac cacgggtgat agaaacagaa      600

acgactgctt cacgcagttc ggttttagca accagttcca gagcgatttc tttaacttcg      660

tcctgggtca gcggcgggat gatacgcata gattcagcgt tagagaacag acgttcgatg      720

tggtcgtcca gacggaaagc gttaccgttc caaacgtgga acccggtgta ggtaacgtca      780

gagtgcaggt aaccctggtc gaagatagag atacgagctt cagacggcgg aacgaaagca      840

ccttcgatcc aagcagcacc accagccagc gggttagccg ggtccagttc gtagtcagag      900

taggtgatat agtccagacc ggtgtcgtgg gtgtaaacga tttcagaggt gtcagcagag      960

aaagccat                                                               968


<210>  5
<211>  342
<212>  PRT
<213>  artificial

<220>
<223>  the amino acid sequence of Arthrobacter-derived transaminase mutant 3

<400>  5

Ala Ala Ala Ile Thr Ile Ile Thr Thr Ala Arg Ile Arg Tyr Cys Thr 
1               5                   10                  15      


Gly Val Ser Arg Glu Glu Asp Ser Thr Phe Ser Ser Gln Gly Arg Arg 
            20                  25                  30          


Met Ile Asp Trp Val Thr Gly Pro Gly Thr Pro Ser Glu Ile Gly Leu 
        35                  40                  45              


Pro Ser Thr Glu Thr Asn Gly Gln Thr Pro Pro Pro Val Glu Gln Pro 
    50                  55                  60                  


Arg Thr Ser Ser Ala Ser Ser Ser Ser Ala Arg Val Met Ser Ala Arg 
65                  70                  75                  80  


Ile Ala Ser Gly Pro Arg Asp Ser Ala Ile Ser Arg Thr Val Leu Arg 
                85                  90                  95      


Val Ile Pro Gly Arg Ala Ala Arg Pro Gly Glu Arg Thr Thr Pro Ser 
            100                 105                 110         


Leu Ile Thr Thr Thr Leu Lys Pro Gly Pro Ser Ala Ser Arg Pro Ser 
        115                 120                 125             


Pro Ser Ser Ser Ser Gly Ser Ser Lys Pro Arg Ser Trp Val Ser Gly 
    130                 135                 140                 


Ile Ala Arg Ile Arg Ser Pro His Trp Lys Phe Leu Thr Cys Gly Ser 
145                 150                 155                 160 


Ile Glu Glu Arg Gly Val Arg Arg Thr Asp Gly Ala Thr Ile Ala Gly 
                165                 170                 175     


Thr Pro Ser Arg Met Arg Ser Asn Gly Thr Ile His Trp Tyr Gly Thr 
            180                 185                 190         


Ala Tyr Met Gly Thr Cys Gly Arg Cys Leu Val Met Ser Arg Ile Tyr 
        195                 200                 205             


Gly Val Glu Glu Gly Pro Arg Val Ile Glu Thr Glu Thr Ile Ala Ser 
    210                 215                 220                 


Arg Ser Ser Val Leu Ala Thr Ser Ser Arg Ala Ile Ser Leu Thr Ser 
225                 230                 235                 240 


Ser Trp Val Ser Gly Gly Met Ile Arg Ile Asp Ser Ala Leu Glu Asn 
                245                 250                 255     


Arg Arg Ser Met Trp Ser Ser Arg Arg Lys Ala Leu Pro Phe Gln Thr 
            260                 265                 270         


Trp Asn Pro Val Gly Val Ala Ser Glu Val Arg Gly Pro Trp Ser Lys 
        275                 280                 285             


Ile Glu Ile Arg Ala Ser Asp Gly Gly Thr Lys Ala Pro Ser Ile Gln 
    290                 295                 300                 


Ala Ala Pro Pro Ala Ser Gly Leu Ala Gly Ser Ser Ser Trp Ser Glu 
305                 310                 315                 320 


Gly Val Ile Gly Ser Arg Pro Val Ser Trp Val Gly Thr Ile Ser Glu 
                325                 330                 335     


Val Ser Ala Glu Lys Ala 
            340         


<210>  6
<211>  1028
<212>  DNA
<213>  artificial

<220>
<223>  the gene sequence of Arthrobacter-derived transaminase mutant 3

<400>  6
gcagcagcca tcaccatcat caccacagcc aggatccggt actgtaccgg ggtcagcaga       60

gaagaagatt caacgttcag ttcccagtaa cgacggatga tagactgggt aaccggaccc      120

ggaacaccgt cagagatcgg gttaccgtca acagaaacga acggccaaac accaccaccg      180

gttgagcaac ccagaacttc gtcagcgtcc agcagttcag ccagggtgat gtcagccagg      240

atagcttcgt gacccagaga ttcagcgatt tccagaacgg ttttacgggt gatacccggc      300

agagcagcac gacccggaga acgaacaaca ccgtctttga taacaacaac gttgaaaccc      360

ggaccttcag ccagcagacc gtcaccgtcc agcagcagcg gcagctcgaa accacggtcg      420

tgggtttcct gaattgcacg gatcaggtca ccccactgga agtttttaac ctgcgggtcg      480

atagaagaac gcggagtacg acgaacagac tgagcaacca tagcgtgaac accgtcacgg      540

atgcggtcaa acggtacgat ccactggtac ggaacagcgt acatgtaaac ctgcggacga      600

tgtttggtga tgtcacgaat atatggggta gaagagtaac cacgggtgat agaaacagaa      660

acgattgctt cacgcagttc ggttttagca accagttcca gagcgatttc tttaacttcg      720

tcctgggtca gcggcgggat gatacgcata gattcagcgt tagagaacag acgttcgatg      780

tggtcgtcca gacggaaagc gttaccgttc caaacgtgga acccggtgta ggtagcgtca      840

gaggtcaggt aaccctggtc gaagatagag atacgagctt cagacggcgg aacgaaagca      900

ccttcgatcc aagcagcacc accagccagc gggttagccg ggtccagttc gtggtcagag      960

taggtgatat agtccagacc ggtgtcgtgg gtgtaaacga tttcagaggt gtcagcagag     1020

aaagccat                                                              1028


<210>  7
<211>  342
<212>  PRT
<213>  artificial

<220>
<223>  the amino acid sequence of Arthrobacter-derived transaminase mutant 4

<400>  7

Ala Ala Ala Ile Thr Ile Ile Thr Thr Ala Arg Ile Arg Tyr Cys Thr 
1               5                   10                  15      


Gly Val Ser Arg Glu Glu Asp Ser Thr Phe Ser Ser Gln Gly Arg Arg 
            20                  25                  30          


Met Ile Asp Trp Val Thr Gly Pro Gly Thr Pro Ser Glu Ile Gly Leu 
        35                  40                  45              


Pro Ser Thr Glu Thr Asn Gly Gln Thr Pro Pro Pro Val Glu Gln Pro 
    50                  55                  60                  


Arg Thr Ser Ser Ala Ser Ser Ser Ser Ala Arg Val Met Ser Ala Arg 
65                  70                  75                  80  


Ile Ala Ser Gly Pro Arg Asp Ser Ala Ile Ser Arg Thr Val Leu Arg 
                85                  90                  95      


Val Ile Pro Gly Arg Ala Ala Arg Pro Gly Glu Arg Thr Thr Pro Ser 
            100                 105                 110         


Leu Ile Thr Thr Thr Leu Lys Pro Gly Pro Ser Ala Ser Arg Pro Ser 
        115                 120                 125             


Gln Ser Ser Ser Ser Gly Ser Ser Lys Pro Arg Ser Trp Val Ser Gly 
    130                 135                 140                 


Ile Ala Arg Ile Arg Ser Pro His Trp Lys Phe Leu Thr Cys Gly Ser 
145                 150                 155                 160 


Ile Glu Glu Arg Gly Val Arg Arg Thr Asp Gly Ala Thr Ile Ala Gly 
                165                 170                 175     


Thr Pro Ser Arg Met Arg Ser Asn Gly Thr Ile His Trp Tyr Gly Thr 
            180                 185                 190         


Ala Tyr Met Gly Thr Cys Gly Arg Cys Leu Val Met Ser Arg Ser Asn 
        195                 200                 205             


Gly Val Glu Glu Gly Pro Arg Val Ile Glu Thr Glu Thr Ile Ala Ser 
    210                 215                 220                 


Arg Ser Ser Val Leu Ala Thr Ser Ser Arg Ala Ile Ser Leu Thr Ser 
225                 230                 235                 240 


Ser Trp Val Ser Gly Gly Met Ile Arg Ile Asp Ser Ala Leu Glu Asn 
                245                 250                 255     


Arg Arg Ser Met Trp Ser Ser Arg Arg Lys Ala Leu Pro Phe Gln Thr 
            260                 265                 270         


Trp Lys Val Val Gly Val Ala Ser Glu Val Gly Gly Pro Trp Ser Lys 
        275                 280                 285             


Ile Glu Ile Arg Ala Ser Asp Gly Gly Thr Lys Ala Pro Ser Ile Gln 
    290                 295                 300                 


Ala Ala Pro Pro Ala Ser Gly Leu Ala Gly Ser Ser Ser Gly Ser Glu 
305                 310                 315                 320 


Gly Val Ile Gly Ser Arg Pro Val Ser Trp Val Gly Thr Ile Ser Glu 
                325                 330                 335     


Val Ser Ala Glu Lys Ala 
            340         


<210>  8
<211>  1028
<212>  DNA
<213>  artificial

<220>
<223>  the gene sequence of Arthrobacter-derived transaminase mutant 4

<400>  8
gcagcagcca tcaccatcat caccacagcc aggatccggt actgtaccgg ggtcagcaga       60

gaagaagatt caacgttcag ttcccagtaa cgacggatga tagactgggt aaccggaccc      120

ggaacaccgt cagagatcgg gttaccgtca acagaaacga acggccaaac accaccaccg      180

gttgagcaac ccagaacttc gtcagcgtcc agcagttcag ccagggtgat gtcagccagg      240

atagcttcgt gacccagaga ttcagcgatt tccagaacgg ttttacgggt gatacccggc      300

agagcagcac gacccggaga acgaacaaca ccgtctttga taacaacaac gttgaaaccc      360

ggaccttcag ccagcagacc gtcgcagtcc agcagcagcg gcagctcgaa accacggtcg      420

tgggtttcct gaattgcacg gatcaggtca ccccactgga agtttttaac ctgcgggtcg      480

atagaagaac gcggagtacg acgaacagac tgagcaacca tagcgtgaac accgtcacgg      540

atgcggtcaa acggtacgat ccactggtac ggaacagcgt acatgtaaac ctgcggacga      600

tgtttggtga tgtcacgctc gaatggggta gaagagtaac cacgggtgat agaaacagaa      660

acgattgctt cacgcagttc ggttttagca accagttcca gagcgatctc tttaacttcg      720

tcctgggtca gcggcgggat gatacggata gattccgcgt tagagaacag acgttcgatg      780

tggtcgtcca gacggaaagc gttaccgttc caaacgtgga aggtggtgta ggtagcgtca      840

gaagtatagt aaccctggtc gaagatagag atacgagctt cagacggcgg aacgaaagca      900

ccttcgatcc aagcagcacc accagccagc gggttagccg ggtccagttc gtagtcagag      960

taggtgatat agtccagacc ggtgtcgtgg gtgtaaacga tttcagaggt gtcagctgag     1020

aaagccat                                                              1028


