                         SEQUENCE LISTING

<110>  Novozymes A/S
       Novozymes North America, Inc.
       Borchert, Martin
       Saunders, Paria
 
<120>  Heat-stable Caminibacter carbonic anhydrases and their use

<130>  11590-WO-PCT

<150>  US 61/220,636
<151>  2009-06-26

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  675
<212>  DNA
<213>  Caminibacter mediatlanticus

<400>  1
acttggagtt atagtggaaa aactggacct gagtattggg gagatttaaa aaaagaatat      60

caaatgtgta aaattgggaa aaatcaatca ccaattgata tcaaaacaaa ttcaactcaa     120

acatttaata caaacttaaa accatttaaa attaaatatt taggtaaagg ttatgaagta     180

ataaacaatg gtcatacaat taaagtaaaa acagaaggca aaaactgtgt tagaattgat     240

ggtattaaat ttaaattagc acaacttcac tttcatacac caagtgaaaa tacaataaat     300

ggaaaacatt ttccaatgga agcacactat gtacatttag ataaaaatgg aaacattaca     360

gttttagcgg ttatgtataa aataggtaaa gaaaataaat cattaaataa aatgttagct     420

gttttaccaa caaaagttgg tgaagaaaat aaagttatgg gtaatctaaa tccaatggaa     480

ttattaccaa aaaataaagc atattataga tttaatggtt cattaacaac tccaccttgt     540

agtgaaggtg ttagatggat agtatttaaa acaccagttg agatttcaca agctcaatat     600

gaaaaaatgc acgctgtaat gggtactaac aatagaccag tccagccaat taatgcaagg     660

gttattttaa aataa                                                      675


<210>  2
<211>  259
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fusion of Savinase signal peptide and CA from C.mediatlanicus

<400>  2

Met Lys Lys Pro Leu Gly Lys Ile Val Ala Ser Thr Ala Leu Leu Ile 
1               5                   10                  15      


Ser Val Ala Phe Ser Ser Ser Ile Ala Ser Ala Ala Ser Ser Tyr Asn 
            20                  25                  30          


Tyr His Ala Thr Trp Ser Tyr Ser Gly Lys Thr Gly Pro Glu Tyr Trp 
        35                  40                  45              


Gly Asp Leu Lys Lys Glu Tyr Gln Met Cys Lys Ile Gly Lys Asn Gln 
    50                  55                  60                  


Ser Pro Ile Asp Ile Lys Thr Asn Ser Thr Gln Thr Phe Asn Thr Asn 
65                  70                  75                  80  


Leu Lys Pro Phe Lys Ile Lys Tyr Leu Gly Lys Gly Tyr Glu Val Ile 
                85                  90                  95      


Asn Asn Gly His Thr Ile Lys Val Lys Thr Glu Gly Lys Asn Cys Val 
            100                 105                 110         


Arg Ile Asp Gly Ile Lys Phe Lys Leu Ala Gln Leu His Phe His Thr 
        115                 120                 125             


Pro Ser Glu Asn Thr Ile Asn Gly Lys His Phe Pro Met Glu Ala His 
    130                 135                 140                 


Tyr Val His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu Ala Val Met 
145                 150                 155                 160 


Tyr Lys Ile Gly Lys Glu Asn Lys Ser Leu Asn Lys Met Leu Ala Val 
                165                 170                 175     


Leu Pro Thr Lys Val Gly Glu Glu Asn Lys Val Met Gly Asn Leu Asn 
            180                 185                 190         


Pro Met Glu Leu Leu Pro Lys Asn Lys Ala Tyr Tyr Arg Phe Asn Gly 
        195                 200                 205             


Ser Leu Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp Ile Val Phe 
    210                 215                 220                 


Lys Thr Pro Val Glu Ile Ser Gln Ala Gln Tyr Glu Lys Met His Ala 
225                 230                 235                 240 


Val Met Gly Thr Asn Asn Arg Pro Val Gln Pro Ile Asn Ala Arg Val 
                245                 250                 255     


Ile Leu Lys 
            


<210>  3
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Polypeptide with savinase signal peptide, HQ tag, Throbin 
       cleavage site and CA from C.mediatlanicus

<400>  3

Met Lys Lys Pro Leu Gly Lys Ile Val Ala Ser Thr Ala Leu Leu Ile 
1               5                   10                  15      


Ser Val Ala Phe Ser Ser Ser Ile Ala Ser Ala His Gln His Gln His 
            20                  25                  30          


Gln His Pro Arg Thr Trp Ser Tyr Ser Gly Lys Thr Gly Pro Glu Tyr 
        35                  40                  45              


Trp Gly Asp Leu Lys Lys Glu Tyr Gln Met Cys Lys Ile Gly Lys Asn 
    50                  55                  60                  


Gln Ser Pro Ile Asp Ile Lys Thr Asn Ser Thr Gln Thr Phe Asn Thr 
65                  70                  75                  80  


Asn Leu Lys Pro Phe Lys Ile Lys Tyr Leu Gly Lys Gly Tyr Glu Val 
                85                  90                  95      


Ile Asn Asn Gly His Thr Ile Lys Val Lys Thr Glu Gly Lys Asn Cys 
            100                 105                 110         


Val Arg Ile Asp Gly Ile Lys Phe Lys Leu Ala Gln Leu His Phe His 
        115                 120                 125             


Thr Pro Ser Glu Asn Thr Ile Asn Gly Lys His Phe Pro Met Glu Ala 
    130                 135                 140                 


His Tyr Val His Leu Asp Lys Asn Gly Asn Ile Thr Val Leu Ala Val 
145                 150                 155                 160 


Met Tyr Lys Ile Gly Lys Glu Asn Lys Ser Leu Asn Lys Met Leu Ala 
                165                 170                 175     


Val Leu Pro Thr Lys Val Gly Glu Glu Asn Lys Val Met Gly Asn Leu 
            180                 185                 190         


Asn Pro Met Glu Leu Leu Pro Lys Asn Lys Ala Tyr Tyr Arg Phe Asn 
        195                 200                 205             


Gly Ser Leu Thr Thr Pro Pro Cys Ser Glu Gly Val Arg Trp Ile Val 
    210                 215                 220                 


Phe Lys Thr Pro Val Glu Ile Ser Gln Ala Gln Tyr Glu Lys Met His 
225                 230                 235                 240 


Ala Val Met Gly Thr Asn Asn Arg Pro Val Gln Pro Ile Asn Ala Arg 
                245                 250                 255     


Val Ile Leu Lys 
            260 


<210>  4
<211>  47
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  4
tttagttcat cgatcgcatc ggctgcgtct tcttacaact accacgc                    47


<210>  5
<211>  47
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  5
gccaaggccg gttttttatg ttttacttaa ggattacgcg agcattg                    47


<210>  6
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  6
catcagcacc aacaccagca tcctaggact tggtcttact ctggcaag                   48


<210>  7
<211>  241
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alpha-amylase signal peptide fused to Methanosarcina thermophila 
       TM-1 carbonic anhydrase

<400>  7

Met Lys Gln Gln Lys Arg Leu Tyr Ala Arg Leu Leu Thr Leu Leu Phe 
1               5                   10                  15      


Ala Leu Ile Phe Leu Leu Pro His Ser Ala Ala Ala Gln Glu Ile Thr 
            20                  25                  30          


Val Asp Glu Phe Ser Asn Ile Arg Glu Asn Pro Val Thr Pro Trp Asn 
        35                  40                  45              


Pro Glu Pro Ser Ala Pro Val Ile Asp Pro Thr Ala Tyr Ile Asp Pro 
    50                  55                  60                  


Gln Ala Ser Val Ile Gly Glu Val Thr Ile Gly Ala Asn Val Met Val 
65                  70                  75                  80  


Ser Pro Met Ala Ser Ile Arg Ser Asp Glu Gly Met Pro Ile Phe Val 
                85                  90                  95      


Gly Asp Arg Ser Asn Val Gln Asp Gly Val Val Leu His Ala Leu Glu 
            100                 105                 110         


Thr Ile Asn Glu Glu Gly Glu Pro Ile Glu Asp Asn Ile Val Glu Val 
        115                 120                 125             


Asp Gly Lys Glu Tyr Ala Val Tyr Ile Gly Asn Asn Val Ser Leu Ala 
    130                 135                 140                 


His Gln Ser Gln Val His Gly Pro Ala Ala Val Gly Asp Asp Thr Phe 
145                 150                 155                 160 


Ile Gly Met Gln Ala Phe Val Phe Lys Ser Lys Val Gly Asn Asn Cys 
                165                 170                 175     


Val Leu Glu Pro Arg Ser Ala Ala Ile Gly Val Thr Ile Pro Asp Gly 
            180                 185                 190         


Arg Tyr Ile Pro Ala Gly Met Val Val Thr Ser Gln Ala Glu Ala Asp 
        195                 200                 205             


Lys Leu Pro Glu Val Thr Asp Asp Tyr Ala Tyr Ser His Thr Asn Glu 
    210                 215                 220                 


Ala Val Val Tyr Val Asn Val His Leu Ala Glu Gly Tyr Lys Glu Thr 
225                 230                 235                 240 


Ser 
    


<210>  8
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  8
cttgctgcct cattctgcag ccgcgcaaga gatcactgtt ga                         42


<210>  9
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  9
tccgatcccc ttttccattc tactttatga agtctctttg tagc                       44


<210>  10
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alignment example

<400>  10

Ala Cys Met Ser His Thr Trp Gly Glu Arg Asn Leu 
1               5                   10          


<210>  11
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alignment example

<400>  11

His Gly Trp Gly Glu Asp Ala Asn Leu Ala Met Asn Pro Ser 
1               5                   10                  


<210>  12
<211>  732
<212>  DNA
<213>  Caminibacter hydrogeniphilus

<400>  12
atgaaaaaaa tattaacaat cgcggcactt gcttcagcaa tgttcgcaag caactacgca      60

catggcacat ggagctacag cggaaaaaca ggtcctgaat actggggaga cttaaaaccg     120

gaatataaaa tgtgtaaaat aggaaaaaat caatctccta ttgatatcag tggagctgta     180

agcgctactc ttacacctgt aaatatttac tatgacgtaa aagcaaaaac atttctaaac     240

aacggccata ctttaaaagc tgaaatgaaa gacggagcaa aactctatat agacggaaaa     300

gaatttagac tgcttcagtt ccatttccat accccaagcg aaaacacaat aaacggtgaa     360

tattttccaa tggaaggaca ttttgttcat tctacaaaag acggtgagct agcggtagta     420

tctgtaatgt ttaaaatcgg taaatataat cccgctattc aaaaacttat aaataatatg     480

ccaaaacacg cgggtgagaa gaaaaatatc tgttctgcta atctaaaagc aaaagattta     540

ttgccaaaaa gtttagagta ttacagattt aacggttctt taaccactcc tccttgtact     600

gaaggggtta gatggttcgt attaaaagaa cctgttcaaa tgagcgccaa gcaggtaaaa     660

gaatttgaaa aaattatggg taaaaacaac cgcccaatcc aaccaatcaa cgcaagagta     720

atactaaaat aa                                                         732


<210>  13
<211>  243
<212>  PRT
<213>  Caminibacter hydrogeniphilus

<400>  13

Met Lys Lys Ile Leu Thr Ile Ala Ala Leu Ala Ser Ala Met Phe Ala 
1               5                   10                  15      


Ser Asn Tyr Ala His Gly Thr Trp Ser Tyr Ser Gly Lys Thr Gly Pro 
            20                  25                  30          


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


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


Thr Pro Val Asn Ile Tyr Tyr Asp Val Lys Ala Lys Thr Phe Leu Asn 
65                  70                  75                  80  


Asn Gly His Thr Leu Lys Ala Glu Met Lys Asp Gly Ala Lys Leu Tyr 
                85                  90                  95      


Ile Asp Gly Lys Glu Phe Arg Leu Leu Gln Phe His Phe His Thr Pro 
            100                 105                 110         


Ser Glu Asn Thr Ile Asn Gly Glu Tyr Phe Pro Met Glu Gly His Phe 
        115                 120                 125             


Val His Ser Thr Lys Asp Gly Glu Leu Ala Val Val Ser Val Met Phe 
    130                 135                 140                 


Lys Ile Gly Lys Tyr Asn Pro Ala Ile Gln Lys Leu Ile Asn Asn Met 
145                 150                 155                 160 


Pro Lys His Ala Gly Glu Lys Lys Asn Ile Cys Ser Ala Asn Leu Lys 
                165                 170                 175     


Ala Lys Asp Leu Leu Pro Lys Ser Leu Glu Tyr Tyr Arg Phe Asn Gly 
            180                 185                 190         


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


Lys Glu Pro Val Gln Met Ser Ala Lys Gln Val Lys Glu Phe Glu Lys 
    210                 215                 220                 


Ile Met Gly Lys Asn Asn Arg Pro Ile Gln Pro Ile Asn Ala Arg Val 
225                 230                 235                 240 


Ile Leu Lys 
            


<210>  14
<211>  774
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of Savinase signal peptide and CA from C.hydrogeniphilus

<400>  14
atgaagaaac cgttggggaa aattgtcgca agcaccgcac tactcatttc tgttgctttt      60

agttcatcga tcgcatcggc tcatcagcac caacaccagc atcctaggac atggagctac     120

agcggaaaaa caggtcctga atactgggga gacttaaaac cggaatataa aatgtgtaaa     180

ataggaaaaa atcaatctcc tattgatatc agtggagctg taagcgctac tcttacacct     240

gtaaatattt actatgacgt aaaagcaaaa acatttctaa acaacggcca tactttaaaa     300

gctgaaatga aagacggagc aaaactctat atagacggaa aagaatttag actgcttcag     360

ttccatttcc ataccccaag cgaaaacaca ataaacggtg aatattttcc aatggaagga     420

cattttgttc attctacaaa agacggtgag ctagcggtag tatctgtaat gtttaaaatc     480

ggtaaatata atcccgctat tcaaaaactt ataaataata tgccaaaaca cgcgggtgag     540

aagaaaaata tctgttctgc taatctaaaa gcaaaagatt tattgccaaa aagtttagag     600

tattacagat ttaacggttc tttaaccact cctccttgta ctgaaggggt tagatggttc     660

gtattaaaag aacctgttca aatgagcgcc aagcaggtaa aagaatttga aaaaattatg     720

ggtaaaaaca accgcccaat ccaaccaatc aacgcaagag taatactaaa ataa           774


<210>  15
<211>  257
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fusion of Savinase signal peptide and CA from C.hydrogeniphilus

<400>  15

Met Lys Lys Pro Leu Gly Lys Ile Val Ala Ser Thr Ala Leu Leu Ile 
1               5                   10                  15      


Ser Val Ala Phe Ser Ser Ser Ile Ala Ser Ala His Gln His Gln His 
            20                  25                  30          


Gln His Pro Arg Thr Trp Ser Tyr Ser Gly Lys Thr Gly Pro Glu Tyr 
        35                  40                  45              


Trp Gly Asp Leu Lys Pro Glu Tyr Lys Met Cys Lys Ile Gly Lys Asn 
    50                  55                  60                  


Gln Ser Pro Ile Asp Ile Ser Gly Ala Val Ser Ala Thr Leu Thr Pro 
65                  70                  75                  80  


Val Asn Ile Tyr Tyr Asp Val Lys Ala Lys Thr Phe Leu Asn Asn Gly 
                85                  90                  95      


His Thr Leu Lys Ala Glu Met Lys Asp Gly Ala Lys Leu Tyr Ile Asp 
            100                 105                 110         


Gly Lys Glu Phe Arg Leu Leu Gln Phe His Phe His Thr Pro Ser Glu 
        115                 120                 125             


Asn Thr Ile Asn Gly Glu Tyr Phe Pro Met Glu Gly His Phe Val His 
    130                 135                 140                 


Ser Thr Lys Asp Gly Glu Leu Ala Val Val Ser Val Met Phe Lys Ile 
145                 150                 155                 160 


Gly Lys Tyr Asn Pro Ala Ile Gln Lys Leu Ile Asn Asn Met Pro Lys 
                165                 170                 175     


His Ala Gly Glu Lys Lys Asn Ile Cys Ser Ala Asn Leu Lys Ala Lys 
            180                 185                 190         


Asp Leu Leu Pro Lys Ser Leu Glu Tyr Tyr Arg Phe Asn Gly Ser Leu 
        195                 200                 205             


Thr Thr Pro Pro Cys Thr Glu Gly Val Arg Trp Phe Val Leu Lys Glu 
    210                 215                 220                 


Pro Val Gln Met Ser Ala Lys Gln Val Lys Glu Phe Glu Lys Ile Met 
225                 230                 235                 240 


Gly Lys Asn Asn Arg Pro Ile Gln Pro Ile Asn Ala Arg Val Ile Leu 
                245                 250                 255     


Lys 
    


<210>  16
<211>  774
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  codon optimized C.hydrogeniphilus gene

<400>  16
atgaagaaac cgttggggaa aattgtcgca agcaccgcac tactcatttc tgttgctttt      60

agttcatcga tcgcatcggc tcatcagcac caacaccagc atcctaggac atggtcttac     120

tcaggcaaga caggtccgga gtactggggt gaccttaagc ctgagtacaa gatgtgtaag     180

atcggcaaga accagtctcc aatcgacatc tcaggtgcgg tttcagcgac acttacacca     240

gttaacatct actacgacgt aaaggctaag actttcctta acaacggcca cacacttaag     300

gctgagatga aggatggcgc aaagctttac atcgacggca aggagttccg ccttcttcaa     360

ttccatttcc acacaccttc tgagaacact atcaacggcg agtacttccc tatggagggc     420

cacttcgttc actctacaaa ggacggcgag cttgcggtag tttctgttat gttcaagatc     480

ggcaagtaca acccagctat ccaaaagctt atcaacaaca tgcctaagca cgctggcgag     540

aagaagaaca tctgttcagc gaaccttaag gcaaaggacc ttcttccgaa gtcacttgag     600

tactaccgct tcaacggctc acttacaact ccaccatgta cagagggcgt tcgctggttc     660

gttcttaagg agccagtaca aatgtcagcg aagcaagtta aggagttcga gaagatcatg     720

ggcaagaaca atcgcccaat ccaaccaatc aatgctcgcg ttatccttaa gtaa           774


<210>  17
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  17
atacttcttt acaattttct cg                                               22


<210>  18
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  18
agaagagtgg agtgataaaa gg                                                22


<210>  19
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  19
tttctaaaca acggccatac                                                  20


<210>  20
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  20
ccaaggccgg ttttttatgt tttattttag tattactctt gcgtt                      45


<210>  21
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  21
catcagcacc aacaccagca tacatggagc tacagcggaa aaac                       44


