                         SEQUENCE LISTING

<110>  CO2 Solutions Inc.
       VOYER, Normand
       DAIGLE, Richard
       MADORE, Eric
       FRADETTE, Sylvie
 
<120>  VARIANTS OF THERMOVIBRIO AMMONIFICANS CARBONIC ANHYDRASE AND CO2 
       CAPTURE METHODS USING THERMOVIBRIO AMMONIFICANS CARBONIC 
       ANHYDRASE VARIANTS

<130>  000677-0219

<140>  Not yet assigned
<141>  2016-09-02

<150>  US 62/213,941
<151>  2015-09-03

<150>  US 62/323,150
<151>  2016-04-15

<160>  10    

<170>  PatentIn version 3.5

<210>  1
<211>  744
<212>  DNA
<213>  Thermovibrio ammonificans

<400>  1
gtgaagagag tattggttac cctcggggct gttgcagcac ttgcaacggg cgcggttgca       60

ggtggaggag cccactgggg ttattccggc agcatcgggc cggagcactg gggagattta      120

agccccgaat accttatgtg taaaatcggt aagaaccaat cgcccataga tattaacagc      180

gccgatgcgg ttaaggcgtg tcttgctccc gttagcgtct actacgtttc agacgcaaag      240

tacgttgtta acaacggcca cacaattaag gttgttatgg ggggaagggg ttacgtggtt      300

gttgacggta agcgctttta cctgaagcag ttccactttc acgcccccag cgagcacacc      360

gttaacggca agcactaccc ctttgaagcc cacttcgtcc accttgataa aaacgggaac      420

ataacggtcc ttggcgtttt ctttaaggtt gggaaggaaa accccgagct tgagaaggtg      480

tggcgtgtta tgcccgagga gccgggtcag aagagacacc ttaccgcaag aatcgacccg      540

gagaagctct tgcccgagaa cagggactac tacagatact ccggctctct caccacaccg      600

ccctgctcgg aaggggttag gtggattgtg tttaaagagc cggttgagat gtctcgggag      660

cagcttgaga agttcaggaa agttatgggc tttgacaaca acaggccggt tcagcccctt      720

aatgcaagga aggttatgaa gtag                                             744


<210>  2
<211>  247
<212>  PRT
<213>  Thermovibrio ammonificans


<220>
<221>  SIGNAL
<222>  (1)..(20)

<400>  2

Met Lys Arg Val Leu Val Thr Leu Gly Ala Val Ala Ala Leu Ala Thr 
1               5                   10                  15      


Gly Ala Val Ala Gly Gly Gly Ala His Trp Gly Tyr Ser Gly Ser Ile 
            20                  25                  30          


Gly Pro Glu His Trp Gly Asp Leu Ser Pro Glu Tyr Leu Met Cys Lys 
        35                  40                  45              


Ile Gly Lys Asn Gln Ser Pro Ile Asp Ile Asn Ser Ala Asp Ala Val 
    50                  55                  60                  


Lys Ala Cys Leu Ala Pro Val Ser Val Tyr Tyr Val Ser Asp Ala Lys 
65                  70                  75                  80  


Tyr Val Val Asn Asn Gly His Thr Ile Lys Val Val Met Gly Gly Arg 
                85                  90                  95      


Gly Tyr Val Val Val Asp Gly Lys Arg Phe Tyr Leu Lys Gln Phe His 
            100                 105                 110         


Phe His Ala Pro Ser Glu His Thr Val Asn Gly Lys His Tyr Pro Phe 
        115                 120                 125             


Glu Ala His Phe Val His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu 
    130                 135                 140                 


Gly Val Phe Phe Lys Val Gly Lys Glu Asn Pro Glu Leu Glu Lys Val 
145                 150                 155                 160 


Trp Arg Val Met Pro Glu Glu Pro Gly Gln Lys Arg His Leu Thr Ala 
                165                 170                 175     


Arg Ile Asp Pro Glu Lys Leu Leu Pro Glu Asn Arg Asp Tyr Tyr Arg 
            180                 185                 190         


Tyr Ser Gly Ser Leu Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp 
        195                 200                 205             


Ile Val Phe Lys Glu Pro Val Glu Met Ser Arg Glu Gln Leu Glu Lys 
    210                 215                 220                 


Phe Arg Lys Val Met Gly Phe Asp Asn Asn Arg Pro Val Gln Pro Leu 
225                 230                 235                 240 


Asn Ala Arg Lys Val Met Lys 
                245         


<210>  3
<211>  687
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleic acid sequence encoding SEQ ID NO: 4

<400>  3
atgggtggcg gtgcacattg gggttatagc ggttcgattg gtccagaaca ttggggtgac       60

ttgtccccgg agtacctgat gtgtaaaatc ggtaagaatc aatccccgat tgatattaat      120

agcgcggacg cggttaaggc atgcctggca ccagttagcg tctactatgt cagcgatgcc      180

aaatacgttg tgaacaacgg ccataccatt aaagttgtga tgggcggtcg tggttatgtc      240

gtcgttgatg gcaaacgttt ctacctgaaa cagttccact tccacgcgcc gagcgagcac      300

acggttaacg gcaagcacta cccgttcgag gctcactttg tgcacctgga taagaatggt      360

aatatcaccg ttctgggcgt gtttttcaag gttggcaagg aaaatccgga gctggaaaaa      420

gtgtggcgcg ttatgccgga agaaccgggc cagaagcgtc atttgaccgc ccgtatcgac      480

cctgagaagc tgctgccgga aaaccgcgac tattaccgtt attctggtag cctgacgact      540

ccgccgtgca gcgagggtgt ccgttggatc gtctttaaag agccggtgga gatgagccgc      600

gaacaactgg agaaatttcg taaagtgatg ggttttgaca acaaccgtcc ggtgcagccg      660

ctgaatgcgc gcaaagtcat gaagtaa                                          687


<210>  4
<211>  228
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of wild-type TACA, wherein the 20-amino acid 
       N-terminal signal sequence of the wild-type TACA is replaced with
       a methionine residue.

<400>  4

Met Gly Gly Gly Ala His Trp Gly Tyr Ser Gly Ser Ile Gly Pro Glu 
1               5                   10                  15      


His Trp Gly Asp Leu Ser Pro Glu Tyr Leu Met Cys Lys Ile Gly Lys 
            20                  25                  30          


Asn Gln Ser Pro Ile Asp Ile Asn Ser Ala Asp Ala Val Lys Ala Cys 
        35                  40                  45              


Leu Ala Pro Val Ser Val Tyr Tyr Val Ser Asp Ala Lys Tyr Val Val 
    50                  55                  60                  


Asn Asn Gly His Thr Ile Lys Val Val Met Gly Gly Arg Gly Tyr Val 
65                  70                  75                  80  


Val Val Asp Gly Lys Arg Phe Tyr Leu Lys Gln Phe His Phe His Ala 
                85                  90                  95      


Pro Ser Glu His Thr Val Asn Gly Lys His Tyr Pro Phe Glu Ala His 
            100                 105                 110         


Phe Val His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu Gly Val Phe 
        115                 120                 125             


Phe Lys Val Gly Lys Glu Asn Pro Glu Leu Glu Lys Val Trp Arg Val 
    130                 135                 140                 


Met Pro Glu Glu Pro Gly Gln Lys Arg His Leu Thr Ala Arg Ile Asp 
145                 150                 155                 160 


Pro Glu Lys Leu Leu Pro Glu Asn Arg Asp Tyr Tyr Arg Tyr Ser Gly 
                165                 170                 175     


Ser Leu Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp Ile Val Phe 
            180                 185                 190         


Lys Glu Pro Val Glu Met Ser Arg Glu Gln Leu Glu Lys Phe Arg Lys 
        195                 200                 205             


Val Met Gly Phe Asp Asn Asn Arg Pro Val Gln Pro Leu Asn Ala Arg 
    210                 215                 220                 


Lys Val Met Lys 
225             


<210>  5
<211>  681
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  nucleotide sequence encoding SEQ ID NO: 7

<400>  5
atggaacacg aatggggtta tagcggttcg attggtccag aacattgggg tgacttgtcc       60

ccggagtacc tgatgtgtaa aatcggtaag aatcaatccc cgattgatat taatagcgcg      120

gacgcggtta aggcatgcct ggcaccagtt agcgtctact atgtcagcga tgccaaatac      180

gttgtgaaca acggccatac cattaaagtt gtgatgggcg gtcgtggtta tgtcgtcgtt      240

gatggcaaac gtttctacct gaaacagttc cacttccacg cgccgagcga gcacacggtt      300

aacggcaagc actacccgtt cgaggctcac tttgtgcacc tggataagaa tggtaatatc      360

accgttctgg gcgtgttttt caaggttggc aaggaaaatc cggagctgga aaaagtgtgg      420

cgcgttatgc cggaagaacc gggccagaag cgtcatttga ccgcccgtat cgaccctgag      480

aagctgctgc cggaaaaccg cgactattac cgttattctg gtagcctgac gactccgccg      540

tgcagcgagg gtgtccgttg gatcgtcttt aaagagccgg tggagatgag ccgcgaacaa      600

ctggagaaat ttcgtaaagt gatgggtttt gacaacaacc gtccggtgca gccgctgaat      660

gcgcgcaaag tcatgaagta a                                                681


<210>  6
<211>  226
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of an N-terminal truncated wild-type TACA 
       variant displaying higher expression in a bacterial host, wherein
       the first six amino acids of SEQ ID NO: 4 ("MGGGAH") are replaced
       with the four residues "MEHE".

<400>  6

Met Glu His Glu Trp Gly Tyr Ser Gly Ser Ile Gly Pro Glu His Trp 
1               5                   10                  15      


Gly Asp Leu Ser Pro Glu Tyr Leu Met Cys Lys Ile Gly Lys Asn Gln 
            20                  25                  30          


Ser Pro Ile Asp Ile Asn Ser Ala Asp Ala Val Lys Ala Cys Leu Ala 
        35                  40                  45              


Pro Val Ser Val Tyr Tyr Val Ser Asp Ala Lys Tyr Val Val Asn Asn 
    50                  55                  60                  


Gly His Thr Ile Lys Val Val Met Gly Gly Arg Gly Tyr Val Val Val 
65                  70                  75                  80  


Asp Gly Lys Arg Phe Tyr Leu Lys Gln Phe His Phe His Ala Pro Ser 
                85                  90                  95      


Glu His Thr Val Asn Gly Lys His Tyr Pro Phe Glu Ala His Phe Val 
            100                 105                 110         


His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu Gly Val Phe Phe Lys 
        115                 120                 125             


Val Gly Lys Glu Asn Pro Glu Leu Glu Lys Val Trp Arg Val Met Pro 
    130                 135                 140                 


Glu Glu Pro Gly Gln Lys Arg His Leu Thr Ala Arg Ile Asp Pro Glu 
145                 150                 155                 160 


Lys Leu Leu Pro Glu Asn Arg Asp Tyr Tyr Arg Tyr Ser Gly Ser Leu 
                165                 170                 175     


Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp Ile Val Phe Lys Glu 
            180                 185                 190         


Pro Val Glu Met Ser Arg Glu Gln Leu Glu Lys Phe Arg Lys Val Met 
        195                 200                 205             


Gly Phe Asp Asn Asn Arg Pro Val Gln Pro Leu Asn Ala Arg Lys Val 
    210                 215                 220                 


Met Lys 
225     


<210>  7
<211>  226
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of an N-terminal truncated wild-type TACA 
       variant displaying higher expression in a bacterial host, wherein
       the first six amino acids of SEQ ID NO: 4 ("MGGGAH") are replaced
       with the four residues "MEHE"

<400>  7

Met Glu His Glu Trp Gly Tyr Ser Gly Ser Ile Gly Pro Glu His Trp 
1               5                   10                  15      


Gly Asp Leu Ser Pro Glu Tyr Leu Met Cys Lys Ile Gly Lys Asn Gln 
            20                  25                  30          


Ser Pro Ile Asp Ile Asn Ser Ala Asp Ala Val Lys Ala Cys Leu Ala 
        35                  40                  45              


Pro Val Ser Val Tyr Tyr Val Ser Asp Ala Lys Tyr Val Val Asn Asn 
    50                  55                  60                  


Gly His Thr Ile Lys Val Val Met Gly Gly Arg Gly Tyr Val Val Val 
65                  70                  75                  80  


Asp Gly Lys Arg Phe Tyr Leu Lys Gln Phe His Phe His Ala Pro Ser 
                85                  90                  95      


Glu His Thr Val Asn Gly Lys His Tyr Pro Phe Glu Ala His Phe Val 
            100                 105                 110         


His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu Gly Val Phe Phe Lys 
        115                 120                 125             


Val Gly Lys Glu Asn Pro Glu Leu Glu Lys Val Trp Arg Val Met Pro 
    130                 135                 140                 


Glu Glu Pro Gly Gln Lys Arg His Leu Thr Ala Arg Ile Asp Pro Glu 
145                 150                 155                 160 


Lys Leu Leu Pro Glu Asn Arg Asp Tyr Tyr Arg Tyr Ser Gly Ser Leu 
                165                 170                 175     


Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp Ile Val Phe Lys Glu 
            180                 185                 190         


Pro Val Glu Met Ser Arg Glu Gln Leu Glu Lys Phe Arg Lys Val Met 
        195                 200                 205             


Gly Phe Asp Asn Asn Arg Pro Val Gln Pro Leu Asn Ala Arg Lys Val 
    210                 215                 220                 


Met Lys 
225     


<210>  8
<211>  227
<212>  PRT
<213>  Sulfurihydrogenibium sp.

<400>  8

Met Glu His Glu Trp Ser Tyr Glu Gly Glu Lys Gly Pro Glu His Trp 
1               5                   10                  15      


Ala Gln Leu Lys Pro Glu Phe Phe Trp Cys Lys Leu Lys Asn Gln Ser 
            20                  25                  30          


Pro Ile Asn Ile Asp Lys Lys Tyr Lys Val Lys Ala Asn Leu Pro Lys 
        35                  40                  45              


Leu Asn Leu Tyr Tyr Lys Thr Ala Lys Glu Ser Glu Val Val Asn Asn 
    50                  55                  60                  


Gly His Thr Ile Gln Ile Asn Ile Lys Glu Asp Asn Thr Leu Asn Tyr 
65                  70                  75                  80  


Leu Gly Glu Lys Tyr Gln Leu Lys Gln Phe His Phe His Thr Pro Ser 
                85                  90                  95      


Glu His Thr Ile Glu Lys Lys Ser Tyr Pro Leu Glu Ile His Phe Val 
            100                 105                 110         


His Lys Thr Glu Asp Gly Lys Ile Leu Val Val Gly Val Met Ala Lys 
        115                 120                 125             


Leu Gly Lys Thr Asn Lys Glu Leu Asp Lys Ile Leu Asn Val Ala Pro 
    130                 135                 140                 


Ala Glu Glu Gly Glu Lys Ile Leu Asp Lys Asn Leu Asn Leu Asn Asn 
145                 150                 155                 160 


Leu Ile Pro Lys Asp Lys Arg Tyr Met Thr Tyr Ser Gly Ser Leu Thr 
                165                 170                 175     


Thr Pro Pro Cys Thr Glu Gly Val Arg Trp Ile Val Leu Lys Lys Pro 
            180                 185                 190         


Ile Ser Ile Ser Lys Gln Gln Leu Glu Lys Leu Lys Ser Val Met Val 
        195                 200                 205             


Asn Pro Asn Asn Arg Pro Val Gln Glu Ile Asn Ser Arg Trp Ile Ile 
    210                 215                 220                 


Glu Gly Phe 
225         


<210>  9
<211>  227
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SspCA 6M1

<400>  9

Met Glu His Glu Trp Ser Tyr Glu Gly Glu Lys Gly Pro Glu His Trp 
1               5                   10                  15      


Ala Phe Leu Arg Pro Glu Phe Phe Trp Cys Lys Leu Lys Asn Gln Ser 
            20                  25                  30          


Pro Ile Asn Ile Asp Ser Lys Tyr Lys Val Lys Ala Asn Leu Pro Lys 
        35                  40                  45              


Leu Asn Leu Tyr Tyr Lys Thr Ala Leu Glu Ser Glu Val Val Asn Asn 
    50                  55                  60                  


Gly His Thr Ile Gln Ile Asn Ile Lys Glu Asp Asn Thr Leu Asn Tyr 
65                  70                  75                  80  


Leu Cys Glu Lys Tyr Gln Leu Lys Gln Phe His Phe His Thr Pro Ser 
                85                  90                  95      


Glu His Thr Val Glu Lys Lys Ser Tyr Pro Leu Glu Ile His Phe Val 
            100                 105                 110         


His Lys Thr Glu Asp Gly Lys Ile Leu Val Val Gly Val Met Ala Lys 
        115                 120                 125             


Leu Gly Lys Thr Asn Lys Glu Leu Asp Lys Ile Leu Asn Val Ala Pro 
    130                 135                 140                 


Ala Glu Glu Gly Glu Lys Ile Leu Asp Lys Asn Leu Asn Leu Asn Asn 
145                 150                 155                 160 


Leu Ile Pro Lys Asp Lys Arg Tyr Met Thr Tyr Ser Gly Ser Leu Thr 
                165                 170                 175     


Thr Pro Pro Cys Thr Glu Gly Val Arg Trp Ile Val Leu Lys Lys Pro 
            180                 185                 190         


Ile Ser Ile Ser Lys Gln Gln Leu Glu Lys Leu Lys Ser Val Met Val 
        195                 200                 205             


Asn Pro Asn Asn Arg Pro Val Gln Glu Ile Asn Ser Arg Trp Ile Ile 
    210                 215                 220                 


Glu Gly Phe 
225         


<210>  10
<211>  222
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Amino acid sequence of wild-type TACA beginning at the highly 
       conserved tryptophan residue at position 26 of SEQ ID NO: 2, 
       position 7 of SEQ ID NO: 4, or position 5 of SEQ ID NO: 7

<400>  10

Trp Gly Tyr Ser Gly Ser Ile Gly Pro Glu His Trp Gly Asp Leu Ser 
1               5                   10                  15      


Pro Glu Tyr Leu Met Cys Lys Ile Gly Lys Asn Gln Ser Pro Ile Asp 
            20                  25                  30          


Ile Asn Ser Ala Asp Ala Val Lys Ala Cys Leu Ala Pro Val Ser Val 
        35                  40                  45              


Tyr Tyr Val Ser Asp Ala Lys Tyr Val Val Asn Asn Gly His Thr Ile 
    50                  55                  60                  


Lys Val Val Met Gly Gly Arg Gly Tyr Val Val Val Asp Gly Lys Arg 
65                  70                  75                  80  


Phe Tyr Leu Lys Gln Phe His Phe His Ala Pro Ser Glu His Thr Val 
                85                  90                  95      


Asn Gly Lys His Tyr Pro Phe Glu Ala His Phe Val His Leu Asp Lys 
            100                 105                 110         


Asn Gly Asn Ile Thr Val Leu Gly Val Phe Phe Lys Val Gly Lys Glu 
        115                 120                 125             


Asn Pro Glu Leu Glu Lys Val Trp Arg Val Met Pro Glu Glu Pro Gly 
    130                 135                 140                 


Gln Lys Arg His Leu Thr Ala Arg Ile Asp Pro Glu Lys Leu Leu Pro 
145                 150                 155                 160 


Glu Asn Arg Asp Tyr Tyr Arg Tyr Ser Gly Ser Leu Thr Thr Pro Pro 
                165                 170                 175     


Cys Ser Glu Gly Val Arg Trp Ile Val Phe Lys Glu Pro Val Glu Met 
            180                 185                 190         


Ser Arg Glu Gln Leu Glu Lys Phe Arg Lys Val Met Gly Phe Asp Asn 
        195                 200                 205             


Asn Arg Pro Val Gln Pro Leu Asn Ala Arg Lys Val Met Lys 
    210                 215                 220         


