               SEQUENCE LISTING

<110> Universidade de Aveiro

<120> Metodo para a producao de colagenase recombinante para digestao de colagenios

<130> P163.1 WO

<150> PT107276
<151> 2013-11-07

<160> 8

<170> BiSSAP 1.3

<210> 1
<211> 2748
<212> DNA
<213> Aeromonas hydrophila

<400> 1
atgaacaatc tgggtaccag gttgctgctg ctggcagcac cgggcctgtt tgccacttcg      60

gcgctggcca actactcccc ctccgactgg caacagtatc aactcaaagg tgaatccagc     120

cgtcagctgg gagatcgtct caccgaagtg acctacgagc tgagcgcccg cagcggcggc     180

gccccctatc agcagctgag ggtgtatcgc cgcttcgact ggagcgatgc caacctggct     240

gcgctggcag aacagcagtg cggcgagccg cagctcaaga tcgagctggg ctggcagatc     300

cgctacctca gctgtgaaga ggtggtgccg gctggcaagg ccgttccggc cagcagctat     360

gactatggct acggcatgaa gcagggccgc tgggagccac tggcaggcac cccgaccgcg     420

ccgcgtcagg atcgtctgcc gctggcggag cgggtcatcc tgggtcacag cgagcaggaa     480

ctggatcgct gcgagctcaa cgccgaaggc cgctgcgccg agcaggcatg gcagtaccag     540

ccgcaaaact ggcagcagtt gaaggtgctg gaagagaccc ccaatgaacg ggatggccgc     600

ctcgagcaga tcttcttccg cctgcagccc atcgtgggca gccaggccgc caagcaggtg     660

agcgagctgc acgtctggcg ccagtacacc tggttgctgg aacaagccaa gacccagcaa     720

gagtgcgatg aaccccagac ccgtcaggaa ggggacaaga ccatcagcta ccgggtgtgc     780

cgccagaccc tgcccgccgg cagcgaagtg caggtggtct tgaaggacac cggctatcaa     840

tacccggtgg ggggcagcga atggcagacc ctgccggaaa caaccgagtg gcaggagagc     900

cgggtgctca accaccccat cgtgctggcc agcaaggaag agcagctcga ctgtcgccgt     960

gccgacggcc gcgcctgttc cgagccagat ctgccgggca ccgagctgct ggacgccgaa    1020

gcggccaaga tagtgcagga tgccagtggc cagcctgcgc cggtctggca ggagaactat    1080

ggtcacgatg acaccaagct gctggccgtg tcgcgcggca tccagagcct gctggcggcc    1140

aatcagccgg cccatccagc catgaagctg ctgctggaat acgtgcgtgc ccacaactac    1200

cacaactacg gcaagcacaa ggaagacggc ccggccgccg ccgaggcgct ggccgaggcg    1260

ttgacggcgc tgggcgccca tccgctgctc ttcccggagc aggccagcga cgaggttggt    1320

gcagtcatgg gtgcctggag catcgccctg cacggtcagt tcaagagccc agcagtgcag    1380

agccgctttg gcaccctgct cggcgagttc aaccagatgc tggcctacag cacccgccat    1440

gccagcgaga tcaacggtca gcacgcctgg gcaaccggcc tgttcgatct gctcaacttc    1500

ctcgacttcg ccagcgacta cagcgacccc ttcgccaacg acttccgcca acaggacggt    1560

gagctgcgca agcagctgca cgccctcggc atgagcgaac tcgcgttgtg gaagggacgg    1620

gatggcgccg atctgttcct gctcaacaac gtgctggatg cctacacccg cctctaccgg    1680

gtcgcccgct atacccgccc ggacgagctc gacggctatc gcaaactgtt ggatgactcc    1740

gtcatcgcac tggttcgcca ccacaacctg atccccggtg gccagcagag tcaggatctg    1800

ctggaagaca tgtcactgac cctctccacc tactacctga cctacacgga ccgcaccagc    1860

gaggcctgca tcagcggtga cttcgccggg ctctgcaccc ctgtccgggt ggaggacgtg    1920

ctgccgttcg agcacacctg ctcgccgacc ctgcgcctgc gggcccagga cctgaccatg    1980

gatcaggccg aagggatctg ccgtgaactg ggtgccgaag agcagcagtt ccatcaacag    2040

atggagaccg gctggcagcc ggtggcggac gatcacaacg aggcgctgga actggtggtc    2100

ttcaactcct ccgccgactg gaaacgctat ggcagtgctc tgttcggcgg cgtctccacc    2160

gacaacggcg gcatctacct cgaaggggat ccggctcgcc ccggcaacca ggcccgcttc    2220

ttcgcctacg aggcggagtg gaagcgccca gcgttccagg tgtggaacct gcgccacgag    2280

tatgtgcact acctggacgg ccgcttcaac cagtacggca gcttcggcca ctacccgctc    2340

aaccgcacca cctggtggtc ggaagggctg gcggagttca tcgcccacgg ccagtgcttc    2400

gcccgcggtc tggacaacgt cgccggccgt cctgccagtg atcgtccggc cttggccgac    2460

atcctgcacc tggattacga caagggtggc gagatggtct actcctggtc ctacacggtg    2520

caccgcttcc tgaacgaaac cggtcgcggc gccagctggc tggccatggc ccaggccctg    2580

cgcaacccgg atcagcagca ggccatgagc accttcgaag ccgagctgga ccagctgatt    2640

gccaatgaca gcgaggccta ccagcagtgg ctcggccgcg agctgctgcc ctggtgggaa    2700

gccaacaagg actccgacga gtgcaaggcc aacgactcct cccactga                 2748


<210> 2
<211> 29
<212> DNA
<213> Aeromonas hydrophila

<400> 2
gggataatca tgaacaatct gggtaccag                                      29


<210> 3
<211> 26
<212> DNA
<213> Aeromonas hydrophila

<400> 3
ttaagcttgt gggaggagtc gttggc                                         26


<210> 4
<211> 3007
<212> DNA
<213> Aeromonas hydrophila

<400> 4
taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt      60

tgtttaactt taagaaggag atatacatat gaaatacctg ctgccgaccg ctgctgctgg     120

tctgctgctc ctcgctgccc agccggcgat ggccatgaac aatctgggta ccaggttgct     180

gctgctggca gcaccgggcc tgtttgccac ttcggcgctg gccaactact ccccctccga     240

ctggcaacag tatcaactca aaggtgaatc cagccgtcag ctgggagatc gtctcaccga     300

agtgacctac gagctgagcg cccgcagcgg cggcgccccc tatcagcagc tgagggtgta     360

tcgccgcttc gactggagcg atgccaacct ggctgcgctg gcagaacagc agtgcggcga     420

gccgcagctc aagatcgagc tgggctggca gatccgctac ctcagctgtg aagaggtggt     480

gccggctggc aaggccgttc cggccagcag ctatgactat ggctacggca tgaagcaggg     540

ccgctgggag ccactggcag gcaccccgac cgcgccgcgt caggatcgtc tgccgctggc     600

ggagcgggtc atcctgggtc acagcgagca ggaactggat cgctgcgagc tcaacgccga     660

aggccgctgc gccgagcagg catggcagta ccagccgcaa aactggcagc agttgaaggt     720

gctggaagag acccccaatg aacgggatgg ccgcctcgag cagatcttct tccgcctgca     780

gcccatcgtg ggcagccagg ccgccaagca ggtgagcgag ctgcacgtct ggcgccagta     840

cacctggttg ctggaacaag ccaagaccca gcaagagtgc gatgaacccc agacccgtca     900

ggaaggggac aagaccatca gctaccgggt gtgccgccag accctgcccg ccggcagcga     960

agtgcaggtg gtcttgaagg acaccggcta tcaatacccg gtggggggca gcgaatggca    1020

gaccctgccg gaaacaaccg agtggcagga gagccgggtg ctcaaccacc ccatcgtgct    1080

ggccagcaag gaagagcagc tcgactgtcg ccgtgccgac ggccgcgcct gttccgagcc    1140

agatctgccg ggcaccgagc tgctggacgc cgaagcggcc aagatagtgc aggatgccag    1200

tggccagcct gcgccggtct ggcaggagaa ctatggtcac gatgacacca agctgctggc    1260

cgtgtcgcgc ggcatccaga gcctgctggc ggccaatcag ccggcccatc cagccatgaa    1320

gctgctgctg gaatacgtgc gtgcccacaa ctaccacaac tacggcaagc acaaggaaga    1380

cggcccggcc gccgccgagg cgctggccga ggcgttgacg gcgctgggcg cccatccgct    1440

gctcttcccg gagcaggcca gcgacgaggt tggtgcagtc atgggtgcct ggagcatcgc    1500

cctgcacggt cagttcaaga gcccagcagt gcagagccgc tttggcaccc tgctcggcga    1560

gttcaaccag atgctggcct acagcacccg ccatgccagc gagatcaacg gtcagcacgc    1620

ctgggcaacc ggcctgttcg atctgctcaa cttcctcgac ttcgccagcg actacagcga    1680

ccccttcgcc aacgacttcc gccaacagga cggtgagctg cgcaagcagc tgcacgccct    1740

cggcatgagc gaactcgcgt tgtggaaggg acgggatggc gccgatctgt tcctgctcaa    1800

caacgtgctg gatgcctaca cccgcctcta ccgggtcgcc cgctataccc gcccggacga    1860

gctcgacggc tatcgcaaac tgttggatga ctccgtcatc gcactggttc gccaccacaa    1920

cctgatcccc ggtggccagc agagtcagga tctgctggaa gacatgtcac tgaccctctc    1980

cacctactac ctgacctaca cggaccgcac cagcgaggcc tgcatcagcg gtgacttcgc    2040

cgggctctgc acccctgtcc gggtggagga cgtgctgccg ttcgagcaca cctgctcgcc    2100

gaccctgcgc ctgcgggccc aggacctgac catggatcag gccgaaggga tctgccgtga    2160

actgggtgcc gaagagcagc agttccatca acagatggag accggctggc agccggtggc    2220

ggacgatcac aacgaggcgc tggaactggt ggtcttcaac tcctccgccg actggaaacg    2280

ctatggcagt gctctgttcg gcggcgtctc caccgacaac ggcggcatct acctcgaagg    2340

ggatccggct cgccccggca accaggcccg cttcttcgcc tacgaggcgg agtggaagcg    2400

cccagcgttc caggtgtgga acctgcgcca cgagtatgtg cactacctgg acggccgctt    2460

caaccagtac ggcagcttcg gccactaccc gctcaaccgc accacctggt ggtcggaagg    2520

gctggcggag ttcatcgccc acggccagtg cttcgcccgc ggtctggaca acgtcgccgg    2580

ccgtcctgcc agtgatcgtc cggccttggc cgacatcctg cacctggatt acgacaaggg    2640

tggcgagatg gtctactcct ggtcctacac ggtgcaccgc ttcctgaacg aaaccggtcg    2700

cggcgccagc tggctggcca tggcccaggc cctgcgcaac ccggatcagc agcaggccat    2760

gagcaccttc gaagccgagc tggaccagct gattgccaat gacagcgagg cctaccagca    2820

gtggctcggc cgcgagctgc tgccctggtg ggaagccaac aaggactccg acgagtgcaa    2880

ggccaacgac tcctcccaca agcttgcggc cgcactcgag caccaccacc accacctgag    2940

atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat    3000

aactagc                                                              3007


<210> 5
<211> 927
<212> PRT
<213> Aeromonas hydrophila

<400> 5
Met Asn Asn Leu Gly Thr Arg Leu Leu Leu Leu Ala Ala Pro Gly Leu 
1               5                   10                  15      
Phe Ala Thr Ser Ala Leu Ala Asn Tyr Ser Pro Ser Asp Trp Gln Gln 
            20                  25                  30          
Tyr Gln Leu Lys Gly Glu Ser Ser Arg Gln Leu Gly Asp Arg Leu Thr 
        35                  40                  45              
Glu Val Thr Tyr Glu Leu Ser Ala Arg Ser Gly Gly Ala Pro Tyr Gln 
    50                  55                  60                  
Gln Leu Arg Val Tyr Arg Arg Phe Asp Trp Ser Asp Ala Asn Leu Ala 
65                  70                  75                  80  
Ala Leu Ala Glu Gln Gln Cys Gly Glu Pro Gln Leu Lys Ile Glu Leu 
                85                  90                  95      
Gly Trp Gln Ile Arg Tyr Leu Ser Cys Glu Glu Val Val Pro Ala Gly 
            100                 105                 110         
Lys Ala Val Pro Ala Ser Ser Tyr Asp Tyr Gly Tyr Gly Met Lys Gln 
        115                 120                 125             
Gly Arg Trp Glu Pro Leu Ala Gly Thr Pro Thr Ala Pro Arg Gln Asp 
    130                 135                 140                 
Arg Leu Pro Leu Ala Glu Arg Val Ile Leu Gly His Ser Glu Gln Glu 
145                 150                 155                 160 
Leu Asp Arg Cys Glu Leu Asn Ala Glu Gly Arg Cys Ala Glu Gln Ala 
                165                 170                 175     
Trp Gln Tyr Gln Pro Gln Asn Trp Gln Gln Leu Lys Val Leu Glu Glu 
            180                 185                 190         
Thr Pro Asn Glu Arg Asp Gly Arg Leu Glu Gln Ile Phe Phe Arg Leu 
        195                 200                 205             
Gln Pro Ile Val Gly Ser Gln Ala Ala Lys Gln Val Ser Glu Leu His 
    210                 215                 220                 
Val Trp Arg Gln Tyr Thr Trp Leu Leu Glu Gln Ala Lys Thr Gln Gln 
225                 230                 235                 240 
Glu Cys Asp Glu Pro Gln Thr Arg Gln Glu Gly Asp Lys Thr Ile Ser 
                245                 250                 255     
Tyr Arg Val Cys Arg Gln Thr Leu Pro Ala Gly Ser Glu Val Gln Val 
            260                 265                 270         
Val Leu Lys Asp Thr Gly Tyr Gln Tyr Pro Val Gly Gly Ser Glu Trp 
        275                 280                 285             
Gln Thr Leu Pro Glu Thr Thr Glu Trp Gln Glu Ser Arg Val Leu Asn 
    290                 295                 300                 
His Pro Ile Val Leu Ala Ser Lys Glu Glu Gln Leu Asp Cys Arg Arg 
305                 310                 315                 320 
Ala Asp Gly Arg Ala Cys Ser Glu Pro Asp Leu Pro Gly Thr Glu Leu 
                325                 330                 335     
Leu Asp Ala Glu Ala Ala Lys Ile Val Gln Asp Ala Ser Gly Gln Pro 
            340                 345                 350         
Ala Pro Val Trp Gln Glu Asn Tyr Gly His Asp Asp Thr Lys Leu Leu 
        355                 360                 365             
Ala Val Ser Arg Gly Ile Gln Ser Leu Leu Ala Ala Asn Gln Pro Ala 
    370                 375                 380                 
His Pro Ala Met Lys Leu Leu Leu Glu Tyr Val Arg Ala His Asn Tyr 
385                 390                 395                 400 
His Asn Tyr Gly Lys His Lys Glu Asp Gly Pro Ala Ala Ala Glu Ala 
                405                 410                 415     
Leu Ala Glu Ala Leu Thr Ala Leu Gly Ala His Pro Leu Leu Phe Pro 
            420                 425                 430         
Glu Gln Ala Ser Asp Glu Val Gly Ala Val Met Gly Ala Trp Ser Ile 
        435                 440                 445             
Ala Leu His Gly Gln Phe Lys Ser Pro Ala Val Gln Ser Arg Phe Gly 
    450                 455                 460                 
Thr Leu Leu Gly Glu Phe Asn Gln Met Leu Ala Tyr Ser Thr Arg His 
465                 470                 475                 480 
Ala Ser Glu Ile Asn Gly Gln His Ala Trp Ala Thr Gly Leu Phe Asp 
                485                 490                 495     
Leu Leu Asn Phe Leu Asp Phe Ala Ser Asp Tyr Ser Asp Pro Phe Ala 
            500                 505                 510         
Asn Asp Phe Arg Gln Gln Asp Gly Glu Leu Arg Lys Gln Leu His Ala 
        515                 520                 525             
Leu Gly Met Ser Glu Leu Ala Leu Trp Lys Gly Arg Asp Gly Ala Asp 
    530                 535                 540                 
Leu Phe Leu Leu Asn Asn Val Leu Asp Ala Tyr Thr Arg Leu Tyr Arg 
545                 550                 555                 560 
Val Ala Arg Tyr Thr Arg Pro Asp Glu Leu Asp Gly Tyr Arg Lys Leu 
                565                 570                 575     
Leu Asp Asp Ser Val Ile Ala Leu Val Arg His His Asn Leu Ile Pro 
            580                 585                 590         
Gly Gly Gln Gln Ser Gln Asp Leu Leu Glu Asp Met Ser Leu Thr Leu 
        595                 600                 605             
Ser Thr Tyr Tyr Leu Thr Tyr Thr Asp Arg Thr Ser Glu Ala Cys Ile 
    610                 615                 620                 
Ser Gly Asp Phe Ala Gly Leu Cys Thr Pro Val Arg Val Glu Asp Val 
625                 630                 635                 640 
Leu Pro Phe Glu His Thr Cys Ser Pro Thr Leu Arg Leu Arg Ala Gln 
                645                 650                 655     
Asp Leu Thr Met Asp Gln Ala Glu Gly Ile Cys Arg Glu Leu Gly Ala 
            660                 665                 670         
Glu Glu Gln Gln Phe His Gln Gln Met Glu Thr Gly Trp Gln Pro Val 
        675                 680                 685             
Ala Asp Asp His Asn Glu Ala Leu Glu Leu Val Val Phe Asn Ser Ser 
    690                 695                 700                 
Ala Asp Trp Lys Arg Tyr Gly Ser Ala Leu Phe Gly Gly Val Ser Thr 
705                 710                 715                 720 
Asp Asn Gly Gly Ile Tyr Leu Glu Gly Asp Pro Ala Arg Pro Gly Asn 
                725                 730                 735     
Gln Ala Arg Phe Phe Ala Tyr Glu Ala Glu Trp Lys Arg Pro Ala Phe 
            740                 745                 750         
Gln Val Trp Asn Leu Arg His Glu Tyr Val His Tyr Leu Asp Gly Arg 
        755                 760                 765             
Phe Asn Gln Tyr Gly Ser Phe Gly His Tyr Pro Leu Asn Arg Thr Thr 
    770                 775                 780                 
Trp Trp Ser Glu Gly Leu Ala Glu Phe Ile Ala His Gly Gln Cys Phe 
785                 790                 795                 800 
Ala Arg Gly Leu Asp Asn Val Ala Gly Arg Pro Ala Ser Asp Arg Pro 
                805                 810                 815     
Ala Leu Ala Asp Ile Leu His Leu Asp Tyr Asp Lys Gly Gly Glu Met 
            820                 825                 830         
Val Tyr Ser Trp Ser Tyr Thr Val His Arg Phe Leu Asn Glu Thr Gly 
        835                 840                 845             
Arg Gly Ala Ser Trp Leu Ala Met Ala Gln Ala Leu Arg Asn Pro Asp 
    850                 855                 860                 
Gln Gln Gln Ala Met Ser Thr Phe Glu Ala Glu Leu Asp Gln Leu Ile 
865                 870                 875                 880 
Ala Asn Asp Ser Glu Ala Tyr Gln Gln Trp Leu Gly Arg Glu Leu Leu 
                885                 890                 895     
Pro Trp Trp Glu Ala Asn Lys Asp Ser Asp Glu Cys Lys Ala Asn Asp 
            900                 905                 910         
Ser Ser His Lys Leu Ala Ala Ala Leu Glu His His His His His 
        915                 920                 925         

<210> 6
<211> 19
<212> DNA
<213> Escherichia coli

<400> 6
aatacgactc actataggg                                                 19


<210> 7
<211> 19
<212> DNA
<213> Escherichia coli

<400> 7
gctagttatt gctcagcgg                                                 19


<210> 8
<211> 20
<212> DNA
<213> Escherichia coli

<400> 8
taatacgact cactataggg                                                20


