﻿                         序列表

<110>  华东理工大学
       江苏奥赛康药业股份有限公司

<120>  一种慢生根瘤菌单加氧酶及其在制备手性亚砜中的应用

<160>  6

<170>  Patent In version 3.5

<210>  1
<211>  1461
<212>  DNA
<213>  Bradyrhizobium oligotrophicum ECU1212

<400>  1
atgtcaactg agcatgtcga cgtgctgatc gtcggtgccg ggctgtccgg catcgccgcc        60
gcctatcatc tgcagcacaa atgtccgggc aagcgcttcg ccctcctgga ggggcgcggt       120
gcgctcggcg gcacctggga cctgttccgc tatcccggca tccgctcgga cagcgacatg       180
tacacgctcg gctattcgtt caagccgtgg accgatccga aagccatcgc cgacgggccg       240
cagatcctca aatatgtcca ggacaccgcg accgagaacg gcatcgaccg ccacatccgt       300
ttcaatcatc gcgtccgccg cgcctcgtgg tcgagcgcgg acgcacgctg gacggtcgag       360
gccgagcggc agacggcgca gggcacgacg gaaaccgtgt cgatgacatg cggcttcctt       420
ttcatgtgct ccggctacta ccgctatgag aaaggctatc tgccggactt caagggcatg       480
gccgatttca agggccgcat cgtgcatccg caggcctggc ccgccgacct cgactatgcc       540
ggcaaacgcg tcgtcgtgat cggctccggt gcgacggcgg tgacgctggt gccggcaatg       600
gccaagaccg cggcgcacgt gacgatgctg cagcgctcgc cgacctatgt cgtgtcgcgc       660
ccggcgcagg atgcgctcgc caacaagctg cgcgagcacc tcccggctgg tctcgcctat       720
catctgatcc gctggcgcaa cgtgctgttc gggatgtatt tcttccagct cagccgccgc       780
aagccgcagc gggtcaagca gctgatatta gggggcgtgc gcgccgcgct tggccccgac       840
tacgacgtcg ccacccattt cacgccgcgc tacaacccgt gggaccagcg gttgtgcctg       900
gtgccggatg gcgacctgtt caggaccatc cgcgagcagc gggcgtcggt ggtgacagcc       960
gggatcgaca cgttcaccga gcgcggcctg cgcctctccg acggccgcga gctcgaggcc      1020
gaaatcgtgg tgaccgcgac ggggctggtg ctgcaggttc tcggcggcag cgaggtcgtg      1080
gtcgacggcc gcacggtcga ttttgccaag acgctcaact acaagggcat gatgtattcc      1140
gacgtgccca acatggcggc caccctcggc tacacgaact tctcgtggac gctgaaatgc      1200
gatctcacct gcgaatatgt ctgccgtctc ctcaactaca tggatcgcca tggctatcgc      1260
caatgcgtgc cgcacaacga cgacaccacc gtcacgccgc tgccgtcgct gagcttcagc      1320
tccggctatg tgcagcgctc gattgccgac ctgcccaagc aaggctcgaa gcggccgtgg      1380
cggctgtacc agaactacgc gctggatatc gtctcgctgc ggttcggcaa ggtcgatgat      1440
ggggtgatgc ggtattcgtg a                                                1461

<210>  2
<211>  486
<212>  PRT
<213>  Bradyrhizobium oligotrophicum ECU1212

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

<210>  3
<211>  1461
<212>  DNA
<213>  Artificial Sequence

<220>
<221>  variation

<400>  3
atgtcaactg agcatgtcga cgtgctgatc gtcggtgccg ggctgtccgg catcgccgcc        60
gcctatcatc tgcagcacaa atgtccgggc aagcgcttcg ccctcctgga ggggcgcggt       120
gcgctcggcg gcacctggga cctgttccgc tatcccggca tccgctcgga cagcgacatg       180
tacacgctcg gctattcgtt caagccgtgg accgatccga aagccatcgc cgacgggccg       240
cagatcctca aatatgtcca ggacaccgcg accgagaacg gcatcgaccg ccacatccgt       300
ttcaatcatc gcgtccgccg cgcctcgtgg tcgagcgcgg acgcacgctg gacggtcgag       360
gccgagcggc agacggcgca gggcacgacg gaaaccgtgt cgatgacatg cggcttcctt       420
ttcatgtgct ccggctacta ccgctatgag aaaggctatc tgccggactt caagggcatg       480
gccgatttca agggccgcat cgtgcatccg caggcctggc ccgccgacct cgactatgcc       540
ggcaaacgcg tcgtcgtgat cggctccggt gcgacggcgg tgacgctggt gccggcaatg       600
gccaagaccg cggcgcacgt gacgatgctg cagcgctcgc cgacctatgt cgtgtcgcgc       660
ccggcgcagg atgcgctcgc caacaagctg cgcgagcacc tcccggctgg tctcgcctat       720
catctgatcc gctggcgcaa cgtgctgttc gggatgtatt tcttccagct cagccgccgc       780
aagccgcagc gggtcaagca gctgatatta gggggcgtgc gcgccgcgct tggccccgac       840
tacgacgtcg ccacccattt cacgccgcgc tacaacccgt ggtgccagcg gttgtgcctg       900
gtgccggatg gcgacctgtt caggaccatc cgcgagcagc gggcgtcggt ggtgacagcc       960
gggatcgaca cgttcaccga gcgcggcctg cgcctctccg acggccgcga gctcgaggcc      1020
gaaatcgtgg tgaccgcgac ggggctggtg ctgcaggttc tcggcggcat cgaggtcgtg      1080
gtcgacggcc gcacggtcga ttttgccaag acgctcaact acaagggcat gatgtattcc      1140
gacgtgccca acatggcggc caccctcggc tacacgaacg cattggcgac gctgaaatgc      1200
gatctcacct gcgaatatgt ctgccgtctc ctcaactaca tggatcgcca tggctatcgc      1260
caatgcgtgc cgcacaacga cgacaccacc gtcacgccgc tgccgtcgct gagcttcagc      1320
tccggctatg tgcagcgctc gattgccgac ctgcccaagc aaggctcgaa gcggccgtgg      1380
cggctgtacc agaactacgc gctggatatc gtctcgctgc ggttcggcaa ggtcgatgat      1440
ggggtgatgc ggtattcgtg a                                                1461

<210>  4
<211>  486
<212>  PRT
<213>  Artificial Sequence

<220>
<221>  VARIANT

<400>  4
Met Ser Thr Glu His Val Asp Val Leu Ile Val Gly Ala Gly Leu Ser 
1               5                   10                  15      
Gly Ile Ala Ala Ala Tyr His Leu Gln His Lys Cys Pro Gly Lys Arg 
            20                  25                  30          
Phe Ala Leu Leu Glu Gly Arg Gly Ala Leu Gly Gly Thr Trp Asp Leu 
        35                  40                  45              
Phe Arg Tyr Pro Gly Ile Arg Ser Asp Ser Asp Met Tyr Thr Leu Gly 
    50                  55                  60                  
Tyr Ser Phe Lys Pro Trp Thr Asp Pro Lys Ala Ile Ala Asp Gly Pro 
65                  70                  75                  80  
Gln Ile Leu Lys Tyr Val Gln Asp Thr Ala Thr Glu Asn Gly Ile Asp 
                85                  90                  95      
Arg His Ile Arg Phe Asn His Arg Val Arg Arg Ala Ser Trp Ser Ser 
            100                 105                 110         
Ala Asp Ala Arg Trp Thr Val Glu Ala Glu Arg Gln Thr Ala Gln Gly 
        115                 120                 125             
Thr Thr Glu Thr Val Ser Met Thr Cys Gly Phe Leu Phe Met Cys Ser 
    130                 135                 140                 
Gly Tyr Tyr Arg Tyr Glu Lys Gly Tyr Leu Pro Asp Phe Lys Gly Met 
145                 150                 155                 160 
Ala Asp Phe Lys Gly Arg Ile Val His Pro Gln Ala Trp Pro Ala Asp 
                165                 170                 175     
Leu Asp Tyr Ala Gly Lys Arg Val Val Val Ile Gly Ser Gly Ala Thr 
            180                 185                 190         
Ala Val Thr Leu Val Pro Ala Met Ala Lys Thr Ala Ala His Val Thr 
        195                 200                 205             
Met Leu Gln Arg Ser Pro Thr Tyr Val Val Ser Arg Pro Ala Gln Asp 
    210                 215                 220                 
Ala Leu Ala Asn Lys Leu Arg Glu His Leu Pro Ala Gly Leu Ala Tyr 
225                 230                 235                 240 
His Leu Ile Arg Trp Arg Asn Val Leu Phe Gly Met Tyr Phe Phe Gln 
                245                 250                 255     
Leu Ser Arg Arg Lys Pro Gln Arg Val Lys Gln Leu Ile Leu Gly Gly 
            260                 265                 270         
Val Arg Ala Ala Leu Gly Pro Asp Tyr Asp Val Ala Thr His Phe Thr 
        275                 280                 285             
Pro Arg Tyr Asn Pro Trp Cys Gln Arg Leu Cys Leu Val Pro Asp Gly 
    290                 295                 300                 
Asp Leu Phe Arg Thr Ile Arg Glu Gln Arg Ala Ser Val Val Thr Ala 
305                 310                 315                 320 
Gly Ile Asp Thr Phe Thr Glu Arg Gly Leu Arg Leu Ser Asp Gly Arg 
                325                 330                 335     
Glu Leu Glu Ala Glu Ile Val Val Thr Ala Thr Gly Leu Val Leu Gln 
            340                 345                 350         
Val Leu Gly Gly Ser Glu Val Val Val Asp Gly Arg Thr Val Asp Phe 
        355                 360                 365             
Ala Lys Thr Leu Asn Tyr Lys Gly Met Met Tyr Ser Asp Val Pro Asn 
    370                 375                 380                 
Met Ala Ala Thr Leu Gly Tyr Thr Asn Phe Leu Ala Thr Leu Lys Cys 
385                 390                 395                 400 
Asp Leu Thr Cys Glu Tyr Val Cys Arg Leu Leu Asn Tyr Met Asp Arg 
                405                 410                 415     
His Gly Tyr Arg Gln Cys Val Pro His Asn Asp Asp Thr Thr Val Thr 
            420                 425                 430         
Pro Leu Pro Ser Leu Ser Phe Ser Ser Gly Tyr Val Gln Arg Ser Ile 
        435                 440                 445             
Ala Asp Leu Pro Lys Gln Gly Ser Lys Arg Pro Trp Arg Leu Tyr Gln 
    450                 455                 460                 
Asn Tyr Ala Leu Asp Ile Val Ser Leu Arg Phe Gly Lys Val Asp Asp 
465                 470                 475                 480 
Gly Val Met Arg Tyr Ser 
                485     

<210>  5
<211>  1461
<212>  DNA
<213>  Artificial Sequence

<220>
<221>  variation

<400>  5
atgtcaactg agcatgtcga cgtgctgatc gtcggtgccg ggctgtccgg catcgccgcc        60
gcctatcatc tgcagcacaa atgtccgggc aagcgcttcg ccctcctgga ggggcgcggt       120
gcgctcggcg gcacctggga cctgttccgc tatcccggca tccgctcgga cagcgacatg       180
tacacgctcg gctattcgtt caagccgtgg accgatccga aagccatcgc cgacgggccg       240
cagatcctca aatatgtcca ggacaccgcg accgagaacg gcatcgaccg ccacatccgt       300
ttcaatcatc gcgtccgccg cgcctcgtgg tcgagcgcgg acgcacgctg gacggtcgag       360
gccgagcggc agacggcgca gggcacgacg gaaaccgtgt cgatgacatg cggcttcctt       420
ttcatgtgct ccggctacta ccgctatgag aaaggctatc tgccggactt caagggcatg       480
gccgatttca agggccgcat cgtgcatccg caggcctggc ccgccgacct cgactatgcc       540
ggcaaacgcg tcgtcgtgat cggctccggt gcgacggcgg tgacgctggt gccggcaatg       600
gccaagaccg cggcgcacgt gacgatgctg cagcgctcgc cgacctatgt cgtgtcgcgc       660
ccggcgcagg atgcgctcgc caacaagctg cgcgagcacc tcccggctgg tctcgcctat       720
catctgatcc gctggcgcaa cgtgctgttc gggatgtatt tcttccagct cagccgccgc       780
aagccgcagc gggtcaagca gctgatatta gggggcgtgc gcgccgcgct tggccccgac       840
tacgacgtcg ccacccattt cacgccgcgc tacaacccgt gggaccagcg gttgtgcctg       900
gtgccggatg gcgacctgtt caggaccatc cgcgagcagc gggcgtcggt ggtgacagcc       960
gggatcgaca cgttcaccga gcgcggcctg cgcctctccg acggccgcga gctcgaggcc      1020
gaaatcgtgg tgaccgcgac ggggctggtg ctgcaggttc tcggcggcat cgaggtcgtg      1080
gtcgacggcc gcacggtcga ttttgccaag acgctcaact acaagggcat gatgtattcc      1140
gacgtgccca acatggcggc caccctcggc tacacgaacg catcgtggac gctgaaatgc      1200
gatctcacct gcgaatatgt ctgccgtctc ctcaactaca tggatcgcca tggctatcgc      1260
caatgcgtgc cgcacaacga cgacaccacc gtcacgccgc tgccgtcgct gagcttcagc      1320
tccggctatg tgcagcgctc gattgccgac ctgcccaagc aaggctcgaa gcggccgtgg      1380
cggctgtacc agaactacgc gctggatatc gtctcgctgc ggttcggcaa ggtcgatgat      1440
ggggtgatgc ggtattcgtg a                                                1461

<210>  6
<211>  486
<212>  PRT
<213>  Artificial Sequence

<220>
<221>  VARIANT

<400>  6
Met Ser Thr Glu His Val Asp Val Leu Ile Val Gly Ala Gly Leu Ser 
1               5                   10                  15      
Gly Ile Ala Ala Ala Tyr His Leu Gln His Lys Cys Pro Gly Lys Arg 
            20                  25                  30          
Phe Ala Leu Leu Glu Gly Arg Gly Ala Leu Gly Gly Thr Trp Asp Leu 
        35                  40                  45              
Phe Arg Tyr Pro Gly Ile Arg Ser Asp Ser Asp Met Tyr Thr Leu Gly 
    50                  55                  60                  
Tyr Ser Phe Lys Pro Trp Thr Asp Pro Lys Ala Ile Ala Asp Gly Pro 
65                  70                  75                  80  
Gln Ile Leu Lys Tyr Val Gln Asp Thr Ala Thr Glu Asn Gly Ile Asp 
                85                  90                  95      
Arg His Ile Arg Phe Asn His Arg Val Arg Arg Ala Ser Trp Ser Ser 
            100                 105                 110         
Ala Asp Ala Arg Trp Thr Val Glu Ala Glu Arg Gln Thr Ala Gln Gly 
        115                 120                 125             
Thr Thr Glu Thr Val Ser Met Thr Cys Gly Phe Leu Phe Met Cys Ser 
    130                 135                 140                 
Gly Tyr Tyr Arg Tyr Glu Lys Gly Tyr Leu Pro Asp Phe Lys Gly Met 
145                 150                 155                 160 
Ala Asp Phe Lys Gly Arg Ile Val His Pro Gln Ala Trp Pro Ala Asp 
                165                 170                 175     
Leu Asp Tyr Ala Gly Lys Arg Val Val Val Ile Gly Ser Gly Ala Thr 
            180                 185                 190         
Ala Val Thr Leu Val Pro Ala Met Ala Lys Thr Ala Ala His Val Thr 
        195                 200                 205             
Met Leu Gln Arg Ser Pro Thr Tyr Val Val Ser Arg Pro Ala Gln Asp 
    210                 215                 220                 
Ala Leu Ala Asn Lys Leu Arg Glu His Leu Pro Ala Gly Leu Ala Tyr 
225                 230                 235                 240 
His Leu Ile Arg Trp Arg Asn Val Leu Phe Gly Met Tyr Phe Phe Gln 
                245                 250                 255     
Leu Ser Arg Arg Lys Pro Gln Arg Val Lys Gln Leu Ile Leu Gly Gly 
            260                 265                 270         
Val Arg Ala Ala Leu Gly Pro Asp Tyr Asp Val Ala Thr His Phe Thr 
        275                 280                 285             
Pro Arg Tyr Asn Pro Trp Asp Gln Arg Leu Cys Leu Val Pro Asp Gly 
    290                 295                 300                 
Asp Leu Phe Arg Thr Ile Arg Glu Gln Arg Ala Ser Val Val Thr Ala 
305                 310                 315                 320 
Gly Ile Asp Thr Phe Thr Glu Arg Gly Leu Arg Leu Ser Asp Gly Arg 
                325                 330                 335     
Glu Leu Glu Ala Glu Ile Val Val Thr Ala Thr Gly Leu Val Leu Gln 
            340                 345                 350         
Val Leu Gly Gly Ile Glu Val Val Val Asp Gly Arg Thr Val Asp Phe 
        355                 360                 365             
Ala Lys Thr Leu Asn Tyr Lys Gly Met Met Tyr Ser Asp Val Pro Asn 
    370                 375                 380                 
Met Ala Ala Thr Leu Gly Tyr Thr Asn Ala Ser Trp Thr Leu Lys Cys 
385                 390                 395                 400 
Asp Leu Thr Cys Glu Tyr Val Cys Arg Leu Leu Asn Tyr Met Asp Arg 
                405                 410                 415     
His Gly Tyr Arg Gln Cys Val Pro His Asn Asp Asp Thr Thr Val Thr 
            420                 425                 430         
Pro Leu Pro Ser Leu Ser Phe Ser Ser Gly Tyr Val Gln Arg Ser Ile 
        435                 440                 445             
Ala Asp Leu Pro Lys Gln Gly Ser Lys Arg Pro Trp Arg Leu Tyr Gln 
    450                 455                 460                 
Asn Tyr Ala Leu Asp Ile Val Ser Leu Arg Phe Gly Lys Val Asp Asp 
465                 470                 475                 480 
Gly Val Met Arg Tyr Ser 
                485     