                         SEQUENCE LISTING

<110>  METABOLIC EXPLORER
       VOELKER, FRANCOIS
       DUMON-SEIGNOVERT, LAURENCE
       MEYNIAL-SALLES, ISABELLE
       SOUCAILLE, PHILIPPE
 
<120>  Mutant glycerol dehydrogenase (GlyDH) for the production of a biochemical
        by fermentation

<130>  D27716

<150>  EP 09166812.9
<151>  2009-07-30

<150>  US 61/229,775
<151>  2009-07-30

<160>  46    

<170>  PatentIn version 3.5

<210>  1
<211>  367
<212>  PRT
<213>  Escherichia coli

<400>  1

Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp 
1               5                   10                  15      


Val Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp 
            20                  25                  30          


Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val 
        35                  40                  45              


Glu Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe 
    50                  55                  60                  


Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala 
65                  70                  75                  80  


Glu Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr 
                85                  90                  95      


Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala 
            100                 105                 110         


Ile Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser 
        115                 120                 125             


Val Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro 
    130                 135                 140                 


Asn Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Thr 
145                 150                 155                 160 


Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp 
                165                 170                 175     


Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly 
            180                 185                 190         


Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn 
        195                 200                 205             


Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His 
    210                 215                 220                 


Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu 
225                 230                 235                 240 


Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val 
                245                 250                 255     


His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly 
            260                 265                 270         


Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala 
        275                 280                 285             


Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly 
    290                 295                 300                 


Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala 
305                 310                 315                 320 


Lys Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile 
                325                 330                 335     


His Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu 
            340                 345                 350         


Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu 
        355                 360                 365         


<210>  2
<211>  367
<212>  PRT
<213>  Escherichia coli

<400>  2

Met Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp 
1               5                   10                  15      


Val Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp 
            20                  25                  30          


Leu Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val 
        35                  40                  45              


Glu Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe 
    50                  55                  60                  


Gly Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala 
65                  70                  75                  80  


Glu Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr 
                85                  90                  95      


Leu Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala 
            100                 105                 110         


Ile Ala Pro Thr Ile Ala Ser Asn Asp Ala Pro Cys Ser Ala Leu Ser 
        115                 120                 125             


Val Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro 
    130                 135                 140                 


Asn Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala 
145                 150                 155                 160 


Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp 
                165                 170                 175     


Phe Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly 
            180                 185                 190         


Gly Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn 
        195                 200                 205             


Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His 
    210                 215                 220                 


Val Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu 
225                 230                 235                 240 


Ser Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val 
                245                 250                 255     


His Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly 
            260                 265                 270         


Glu Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala 
        275                 280                 285             


Pro Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly 
    290                 295                 300                 


Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala 
305                 310                 315                 320 


Lys Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile 
                325                 330                 335     


His Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu 
            340                 345                 350         


Leu Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu Met 
        355                 360                 365         1   


Asp Arg Ile Ile Gln Ser Pro Gly Lys Tyr Ile Gln Gly Ala Asp Val 
            5                   10                  15          


Ile Asn Arg Leu Gly Glu Tyr Leu Lys Pro Leu Ala Glu Arg Trp Leu 
        20                  25                  30              


Val Val Gly Asp Lys Phe Val Leu Gly Phe Ala Gln Ser Thr Val Glu 
    35                  40                  45                  


Lys Ser Phe Lys Asp Ala Gly Leu Val Val Glu Ile Ala Pro Phe Gly 
50                  55                  60                  65  


Gly Glu Cys Ser Gln Asn Glu Ile Asp Arg Leu Arg Gly Ile Ala Glu 
                70                  75                  80      


Thr Ala Gln Cys Gly Ala Ile Leu Gly Ile Gly Gly Gly Lys Thr Leu 
            85                  90                  95          


Asp Thr Ala Lys Ala Leu Ala His Phe Met Gly Val Pro Val Ala Ile 
        100                 105                 110             


Ala Pro Thr Ile Ala Ser Asn Asp Ala Pro Cys Ser Ala Leu Ser Val 
    115                 120                 125                 


Ile Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Leu Leu Pro Asn 
130                 135                 140                 145 


Asn Pro Asn Met Val Ile Val Asp Thr Lys Ile Val Ala Gly Ala Pro 
                150                 155                 160     


Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp Phe 
            165                 170                 175         


Glu Ala Arg Ala Cys Ser Arg Ser Gly Ala Thr Thr Met Ala Gly Gly 
        180                 185                 190             


Lys Cys Thr Gln Ala Ala Leu Ala Leu Ala Glu Leu Cys Tyr Asn Thr 
    195                 200                 205                 


Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Ala Glu Gln His Val 
210                 215                 220                 225 


Val Thr Pro Ala Leu Glu Arg Val Ile Glu Ala Asn Thr Tyr Leu Ser 
                230                 235                 240     


Gly Val Gly Phe Glu Ser Gly Gly Leu Ala Ala Ala His Ala Val His 
            245                 250                 255         


Asn Gly Leu Thr Ala Ile Pro Asp Ala His His Tyr Tyr His Gly Glu 
        260                 265                 270             


Lys Val Ala Phe Gly Thr Leu Thr Gln Leu Val Leu Glu Asn Ala Pro 
    275                 280                 285                 


Val Glu Glu Ile Glu Thr Val Ala Ala Leu Ser His Ala Val Gly Leu 
290                 295                 300                 305 


Pro Ile Thr Leu Ala Gln Leu Asp Ile Lys Glu Asp Val Pro Ala Lys 
                310                 315                 320     


Met Arg Ile Val Ala Glu Ala Ala Cys Ala Glu Gly Glu Thr Ile His 
            325                 330                 335         


Asn Met Pro Gly Gly Ala Thr Pro Asp Gln Val Tyr Ala Ala Leu Leu 
        340                 345                 350             


Val Ala Asp Gln Tyr Gly Gln Arg Phe Leu Gln Glu Trp Glu 
    355                 360                 365         


<210>  3
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  3
caccatggac cgcattattc aatc                                              24


<210>  4
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  4
ttattcccac tcttgcagga aacgc                                             25


<210>  5
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  5
gacaccaaaa tcgtcgctgg cacacctgca cgtctgctag cggcg                       45


<210>  6
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  6
cgccgctagc agacgtgcag gtgtgccagc gacgattttg gtgtc                       45


<210>  7
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  7
gcgatcgcac cgactatcgc ctctaacgat gcaccctgca gcgcattg                    48


<210>  8
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  8
caatgcgctg cagggtgcat cgttagaggc gatagtcggt gcgatcgc                    48


<210>  9
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  9
ccgacagtaa gacgggtaag cctg                                              24


<210>  10
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  10
agcttagtaa agccctcgct ag                                                22


<210>  11
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  11
catgggctag ctacgtatta attaaagatc tcctagggag ctcaccggtt aaaaataaga       60

gttaccttaa atggtaactc ttattttttt aggcgcgcca                            100


<210>  12
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  12
agcttggcgc gcctaaaaaa ataagagtta ccatttaagg taactcttat ttttaaccgg       60

tgagctccct aggagatctt taattaatac gtagctagcc                            100


<210>  13
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  13
gacctaggct ctaaaggagc aattatgg                                          28


<210>  14
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  14
cgagctctta ttcccactct tgcagg                                            26


<210>  15
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  15
ggttcagaat ccaacgcaga agcggtgata tcccgtaaaa ccacaggc                    48


<210>  16
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  16
gcctgtggtt ttacgggata tcaccgcttc tgcgttggat tctgaacc                    48


<210>  17
<211>  105
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  17
gttaactacg gatgtacatt atggaactga cgactcgcac tttacctgcg cggaaacata       60

ttgcgctggt ggcacacgat caggcctggt gatgatggcg ggatc                      105


<210>  18
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  18
gggaaattaa gttaccggta gtgcctgttg catacagtac gtgttgttcc agtaacggtt       60

gatgccgttc cacccagctc atcagcatct gtttgcattc agaagaactc gtcaagaagg      120


<210>  19
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  19
tccagtcgcc gcatttcaac gacgcggtcg atattctgat ccccgattat cagcgttatc       60

tcgcggaccg tctgaagtaa tgtaggctgg agctgcttcg                            100


<210>  20
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  20
tgtggaaata ctgaaaaatc tggatgtgcc ggtggcgaga aaaccgtaag aaacaggtgg       60

cgtttgccac ctgtgcaata catatgaata tcctccttag                            100


<210>  21
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  21
atgcttgctg tcagaagatt aacaagatac acattcctta gtttatcatt atcaatactc       60

g                                                                       61


<210>  22
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  22
atcctcgaaa ctaagttctt ggt                                               23


<210>  23
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  23
aaacaccaga acttagtttc gaactagttt attcccactc tt                          42


<210>  24
<211>  62
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  24
tgaaatataa ataacgttct taatactaac ataactataa actagtatgg accgcattat       60

tc                                                                      62


<210>  25
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  25
cctggatgta tttacccggt gattgaataa tgcggtccat actagtttat agttatgtta       60

gtatta                                                                  66


<210>  26
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  26
gaggcaagct aaacagatct ctagacctag gtacccgccg gcaaattaaa gccttcgagc       60


<210>  27
<211>  62
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  27
gcttgagaag gttttgggac gctcgaaggc tttaatttgc taggtctaga gatctgttta       60

gc                                                                      62


<210>  28
<211>  64
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  28
ctattcgtca tcgatgtcta gctcttcaat catctccggt ccactagtgg atctgatatc       60

acct                                                                    64


<210>  29
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  29
atgcttgctg tcagaagatt                                                   20


<210>  30
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  30
tagtatggac cgcattattc                                                   20


<210>  31
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  31
atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc agctggagct catagcttca       60


<210>  32
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  32
tgcgggttgg ggtgtgcaga ttaaagttgt tcatactagt ggatccacta gttctagaaa       60


<210>  33
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  33
catagcaatc taatctaagt tttctagaac tagtggatcc actagtatga acaactttaa       60


<210>  34
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  34
tgaaatataa ataacgttct taatactaac ataactataa actagtttag cgggcggctt       60


<210>  35
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  35
tgtcagccgc cgtatatacg aagccgcccg ctaaactagt ttatagttat gttagtatta       60


<210>  36
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  36
ctccatgtcg ctggccgggt gacccggcgg ggacgaggca gcaaattaaa gccttcgagc       60


<210>  37
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  37
gcttgagaag gttttgggac gctcgaaggc tttaatttgc tgcctcgtcc ccgccgggtc       60


<210>  38
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  38
ttaagcaagg attttcttaa cttcttcggc gacagcatca cagtatagcg accagcattc       60


<210>  39
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  39
atgtctgccc ctaagaagat c                                                 21


<210>  40
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  40
actagtttag cgggcggctt                                                   20


<210>  41
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  41
atgtcgaaag ctacatataa ggaacgtgct gctactcatc gccagatctg tttagcttgc       60


<210>  42
<211>  40
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  42
gtgcgagtcg tcagttccat aatacgcaaa ccgcctctcc                             40


<210>  43
<211>  40
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  43
ggagaggcgg tttgcgtatt atggaactga cgactcgcac                             40


<210>  44
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  44
ttagttttgc tggccgcatc ttctcaaata tgcttccctt tacttcagac ggtccgcgag       60


<210>  45
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  45
ggaactgacg actcgcac                                                     18


<210>  46
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Oligonucleotide

<400>  46
ttagtttcta gagttgatga                                                   20


