                         SEQUENCE LISTING

<110>  Fidia Farmaceutici S.p.A
 
<120>  New process for the production and purification of the collagenase enzyme 
       from Vibrio Alginolyticus

<130>  1484PCT

<150>  PD2012A000118
<151>  2012-04-18

<160>  3     

<170>  PatentIn version 3.5

<210>  1
<211>  739
<212>  PRT
<213>  Vibrio alginolyticus


<220>
<221>  mat_peptide
<222>  (1)..(739)
<223>  Active core of collagenase from Vibrio alginolyticus

<400>  1

Thr Ala Cys Asp Leu Glu Ala Leu Val Thr Glu Ser Ser Asn Gln Leu 
1               5                   10                  15      


Ile Ser Glu Ile Leu Ser Gln Gly Ala Thr Cys Val Asn Gln Leu Phe 
            20                  25                  30          


Ser Ala Glu Ser Arg Ile Gln Glu Ser Val Phe Ser Ser Asp His Met 
        35                  40                  45              


Tyr Asn Ile Ala Lys His Thr Thr Thr Leu Ala Lys Gly Tyr Thr Gly 
    50                  55                  60                  


Gly Gly Ser Asp Glu Leu Glu Thr Leu Phe Leu Tyr Leu Arg Ala Gly 
65                  70                  75                  80  


Tyr Tyr Ala Glu Phe Tyr Asn Asp Asn Ile Ser Phe Ile Glu Trp Val 
                85                  90                  95      


Thr Pro Ala Val Lys Glu Ser Val Asp Ala Phe Val Asn Thr Ala Ser 
            100                 105                 110         


Phe Tyr Glu Asn Ser Asp Arg His Gly Lys Val Leu Ser Glu Val Ile 
        115                 120                 125             


Ile Thr Met Asp Ser Ala Gly Leu Gln His Ala Tyr Leu Pro Gln Val 
    130                 135                 140                 


Thr Gln Trp Leu Thr Arg Trp Asn Asp Gln Tyr Ala Gln His Trp Tyr 
145                 150                 155                 160 


Met Arg Asn Ala Val Asn Gly Val Phe Thr Ile Leu Phe Gly Gly Gln 
                165                 170                 175     


Trp Asn Glu Gln Phe Val Gln Ile Ile Gly Asn Gln Thr Asp Leu Ala 
            180                 185                 190         


Lys Ala Leu Gly Asp Phe Ala Leu Arg Ala Ser Ser Ile Gly Ala Glu 
        195                 200                 205             


Asp Glu Phe Met Ala Ala Asn Ala Gly Arg Glu Leu Gly Arg Leu Thr 
    210                 215                 220                 


Lys Tyr Thr Gly Asn Ala Ser Ser Val Val Lys Ser Gln Leu Ser Arg 
225                 230                 235                 240 


Ile Phe Glu Gln Tyr Glu Met Tyr Gly Arg Gly Asp Ala Val Trp Leu 
                245                 250                 255     


Ala Ala Ala Asp Thr Ala Ser Tyr Tyr Ala Asp Cys Ser Glu Phe Gly 
            260                 265                 270         


Ile Cys Asn Phe Glu Thr Glu Leu Lys Gly Leu Val Leu Ser Gln Thr 
        275                 280                 285             


Tyr Thr Cys Ser Pro Thr Ile Arg Ile Leu Ser Gln Asn Met Thr Gln 
    290                 295                 300                 


Glu Gln His Ala Ala Ala Cys Ser Lys Met Gly Tyr Glu Glu Gly Tyr 
305                 310                 315                 320 


Phe His Gln Ser Leu Glu Thr Gly Glu Gln Pro Val Lys Asp Asp His 
                325                 330                 335     


Asn Thr Gln Leu Gln Val Asn Ile Phe Asp Ser Ser Thr Asp Tyr Gly 
            340                 345                 350         


Lys Tyr Ala Gly Pro Ile Phe Asp Ile Ser Thr Asp Asn Gly Gly Met 
        355                 360                 365             


Tyr Leu Glu Gly Asp Pro Ser Gln Pro Gly Asn Ile Pro Asn Phe Ile 
    370                 375                 380                 


Ala Tyr Glu Ala Ser Tyr Ala Asn Ala Asp His Phe Val Trp Asn Leu 
385                 390                 395                 400 


Glu His Glu Tyr Val His Tyr Leu Asp Gly Arg Phe Asp Leu Tyr Gly 
                405                 410                 415     


Gly Phe Ser His Pro Thr Glu Lys Ile Val Trp Trp Ser Glu Gly Ile 
            420                 425                 430         


Ala Glu Tyr Val Ala Gln Glu Asn Asp Asn Gln Ala Ala Leu Glu Thr 
        435                 440                 445             


Ile Leu Asp Gly Ser Thr Tyr Thr Leu Ser Glu Ile Phe Glu Thr Thr 
    450                 455                 460                 


Tyr Asp Gly Phe Asp Val Asp Arg Ile Tyr Arg Trp Gly Tyr Leu Ala 
465                 470                 475                 480 


Val Arg Phe Met Phe Glu Asn His Lys Asp Asp Val Asn Gln Met Leu 
                485                 490                 495     


Val Glu Thr Arg Gln Gly Asn Trp Ile Asn Tyr Lys Ala Thr Ile Thr 
            500                 505                 510         


Gln Trp Ala Asn Leu Tyr Gln Ser Glu Phe Glu Gln Trp Gln Gln Thr 
        515                 520                 525             


Leu Val Ser Asn Gly Ala Pro Asn Ala Val Ile Thr Ala Asn Ser Lys 
    530                 535                 540                 


Gly Lys Val Gly Glu Ser Ile Thr Phe Ser Ser Glu Asn Ser Thr Asp 
545                 550                 555                 560 


Pro Asn Gly Lys Ile Val Ser Val Leu Trp Asp Phe Gly Asp Gly Ser 
                565                 570                 575     


Thr Ser Thr Gln Thr Lys Pro Thr His Gln Tyr Gly Ser Glu Gly Glu 
            580                 585                 590         


Tyr Ser Val Ser Leu Ser Val Thr Asp Ser Glu Gly Leu Thr Ala Thr 
        595                 600                 605             


Ala Thr His Thr Val Val Ile Ser Ala Leu Gly Gly Asn Asp Thr Leu 
    610                 615                 620                 


Pro Gln Asp Cys Ala Val Gln Ser Lys Val Ser Gly Gly Arg Leu Thr 
625                 630                 635                 640 


Ala Gly Glu Pro Val Cys Leu Ala Asn Gln Gln Thr Ile Trp Leu Ser 
                645                 650                 655     


Val Pro Ala Val Asn Glu Ser Ser Asn Leu Ala Ile Thr Thr Gly Asn 
            660                 665                 670         


Gly Thr Gly Asn Leu Lys Leu Glu Tyr Ser Asn Ser Gly Trp Pro Asp 
        675                 680                 685             


Asp Thr Asn Leu His Gly Trp Ser Asp Asn Ile Gly Asn Gly Glu Cys 
    690                 695                 700                 


Ile Thr Leu Ser Asn Gln Ser Asn Tyr Trp Gly Tyr Val Lys Val Ser 
705                 710                 715                 720 


Gly Asp Phe Glu Asn Ala Ala Ile Val Val Asp Phe Asp Ala Gln Lys 
                725                 730                 735     


Cys Arg Gln 
            


<210>  2
<211>  814
<212>  PRT
<213>  Vibrio alginolyticus


<220>
<221>  PEPTIDE
<222>  (1)..(814)
<223>  Microbial collagenase EC 3.4.24.3 from Vibrio Alginolyticus

<400>  2

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


Thr Gly Val Phe Ala Leu Ser Glu Pro Val Ser Gln Val Thr Glu Gln 
            20                  25                  30          


His Ala His Ser Ala His Thr His Gly Val Glu Phe Asn Arg Val Glu 
        35                  40                  45              


Tyr Gln Pro Thr Ala Thr Leu Pro Ile Gln Pro Ser Lys Ala Thr Arg 
    50                  55                  60                  


Val Gln Ser Leu Glu Ser Leu Asp Glu Ser Ser Thr Ala Cys Asp Leu 
65                  70                  75                  80  


Glu Ala Leu Val Thr Glu Ser Ser Asn Gln Leu Ile Ser Glu Ile Leu 
                85                  90                  95      


Ser Gln Gly Ala Thr Cys Val Asn Gln Leu Phe Ser Ala Glu Ser Arg 
            100                 105                 110         


Ile Gln Glu Ser Val Phe Ser Ser Asp His Met Tyr Asn Ile Ala Lys 
        115                 120                 125             


His Thr Thr Thr Leu Ala Lys Gly Tyr Thr Gly Gly Gly Ser Asp Glu 
    130                 135                 140                 


Leu Glu Thr Leu Phe Leu Tyr Leu Arg Ala Gly Tyr Tyr Ala Glu Phe 
145                 150                 155                 160 


Tyr Asn Asp Asn Ile Ser Phe Ile Glu Trp Val Thr Pro Ala Val Lys 
                165                 170                 175     


Glu Ser Val Asp Ala Phe Val Asn Thr Ala Ser Phe Tyr Glu Asn Ser 
            180                 185                 190         


Asp Arg His Gly Lys Val Leu Ser Glu Val Ile Ile Thr Met Asp Ser 
        195                 200                 205             


Ala Gly Leu Gln His Ala Tyr Leu Pro Gln Val Thr Gln Trp Leu Thr 
    210                 215                 220                 


Arg Trp Asn Asp Gln Tyr Ala Gln His Trp Tyr Met Arg Asn Ala Val 
225                 230                 235                 240 


Asn Gly Val Phe Thr Ile Leu Phe Gly Gly Gln Trp Asn Glu Gln Phe 
                245                 250                 255     


Val Gln Ile Ile Gly Asn Gln Thr Asp Leu Ala Lys Ala Leu Gly Asp 
            260                 265                 270         


Phe Ala Leu Arg Ala Ser Ser Ile Gly Ala Glu Asp Glu Phe Met Ala 
        275                 280                 285             


Ala Asn Ala Gly Arg Glu Leu Gly Arg Leu Thr Lys Tyr Thr Gly Asn 
    290                 295                 300                 


Ala Ser Ser Val Val Lys Ser Gln Leu Ser Arg Ile Phe Glu Gln Tyr 
305                 310                 315                 320 


Glu Met Tyr Gly Arg Gly Asp Ala Val Trp Leu Ala Ala Ala Asp Thr 
                325                 330                 335     


Ala Ser Tyr Tyr Ala Asp Cys Ser Glu Phe Gly Ile Cys Asn Phe Glu 
            340                 345                 350         


Thr Glu Leu Lys Gly Leu Val Leu Ser Gln Thr Tyr Thr Cys Ser Pro 
        355                 360                 365             


Thr Ile Arg Ile Leu Ser Gln Asn Met Thr Gln Glu Gln His Ala Ala 
    370                 375                 380                 


Ala Cys Ser Lys Met Gly Tyr Glu Glu Gly Tyr Phe His Gln Ser Leu 
385                 390                 395                 400 


Glu Thr Gly Glu Gln Pro Val Lys Asp Asp His Asn Thr Gln Leu Gln 
                405                 410                 415     


Val Asn Ile Phe Asp Ser Ser Thr Asp Tyr Gly Lys Tyr Ala Gly Pro 
            420                 425                 430         


Ile Phe Asp Ile Ser Thr Asp Asn Gly Gly Met Tyr Leu Glu Gly Asp 
        435                 440                 445             


Pro Ser Gln Pro Gly Asn Ile Pro Asn Phe Ile Ala Tyr Glu Ala Ser 
    450                 455                 460                 


Tyr Ala Asn Ala Asp His Phe Val Trp Asn Leu Glu His Glu Tyr Val 
465                 470                 475                 480 


His Tyr Leu Asp Gly Arg Phe Asp Leu Tyr Gly Gly Phe Ser His Pro 
                485                 490                 495     


Thr Glu Lys Ile Val Trp Trp Ser Glu Gly Ile Ala Glu Tyr Val Ala 
            500                 505                 510         


Gln Glu Asn Asp Asn Gln Ala Ala Leu Glu Thr Ile Leu Asp Gly Ser 
        515                 520                 525             


Thr Tyr Thr Leu Ser Glu Ile Phe Glu Thr Thr Tyr Asp Gly Phe Asp 
    530                 535                 540                 


Val Asp Arg Ile Tyr Arg Trp Gly Tyr Leu Ala Val Arg Phe Met Phe 
545                 550                 555                 560 


Glu Asn His Lys Asp Asp Val Asn Gln Met Leu Val Glu Thr Arg Gln 
                565                 570                 575     


Gly Asn Trp Ile Asn Tyr Lys Ala Thr Ile Thr Gln Trp Ala Asn Leu 
            580                 585                 590         


Tyr Gln Ser Glu Phe Glu Gln Trp Gln Gln Thr Leu Val Ser Asn Gly 
        595                 600                 605             


Ala Pro Asn Ala Val Ile Thr Ala Asn Ser Lys Gly Lys Val Gly Glu 
    610                 615                 620                 


Ser Ile Thr Phe Ser Ser Glu Asn Ser Thr Asp Pro Asn Gly Lys Ile 
625                 630                 635                 640 


Val Ser Val Leu Trp Asp Phe Gly Asp Gly Ser Thr Ser Thr Gln Thr 
                645                 650                 655     


Lys Pro Thr His Gln Tyr Gly Ser Glu Gly Glu Tyr Ser Val Ser Leu 
            660                 665                 670         


Ser Val Thr Asp Ser Glu Gly Leu Thr Ala Thr Ala Thr His Thr Val 
        675                 680                 685             


Val Ile Ser Ala Leu Gly Gly Asn Asp Thr Leu Pro Gln Asp Cys Ala 
    690                 695                 700                 


Val Gln Ser Lys Val Ser Gly Gly Arg Leu Thr Ala Gly Glu Pro Val 
705                 710                 715                 720 


Cys Leu Ala Asn Gln Gln Thr Ile Trp Leu Ser Val Pro Ala Val Asn 
                725                 730                 735     


Glu Ser Ser Asn Leu Ala Ile Thr Thr Gly Asn Gly Thr Gly Asn Leu 
            740                 745                 750         


Lys Leu Glu Tyr Ser Asn Ser Gly Trp Pro Asp Asp Thr Asn Leu His 
        755                 760                 765             


Gly Trp Ser Asp Asn Ile Gly Asn Gly Glu Cys Ile Thr Leu Ser Asn 
    770                 775                 780                 


Gln Ser Asn Tyr Trp Gly Tyr Val Lys Val Ser Gly Asp Phe Glu Asn 
785                 790                 795                 800 


Ala Ala Ile Val Val Asp Phe Asp Ala Gln Lys Cys Arg Gln 
                805                 810                 


<210>  3
<211>  15
<212>  PRT
<213>  Vibrio Alginolyticus


<220>
<221>  MISC_FEATURE
<222>  (1)..(15)
<223>  N terminal of the mature chain

<400>  3

Thr Ala Cys Asp Leu Glu Ala Leu Val Thr Glu Ser Ser Asn Gln 
1               5                   10                  15  


