Patent ID: 9738883
Date: 2017-08-22
CPC Classifications: A61K,C12N,C12Y

Claim:
1. A process for the production and purification of collagenase from Vibrio alginolyticus chemovar. lophagus , comprising the following stages: Stage A: Inoculation of Stage B: Clarification of the fermented broth thus obtained by tangential flow ultrafiltration with 100-500 kD Molecular Weight Cut-Off (MWCO) cassettes; Stage C: Dialysis and concentration of the clarified medium obtained in stage B, by tangential flow ultrafiltration with 5-30 kD MWCO cassettes; Stage D: Purification of the solution containing collagenase obtained in Stage C, by anion-exchange resin carrying weak basic groups, at a pH of between 6.9 and 7.4; Stage E: Dialysis and concentration of the fractions with collagenolytic activity collected in Stage D, by tangential flow ultrafiltration with 10-50 kD MWCO cassettes; Stage F: Purification of the solution thus obtained, by anion-exchange resin carrying strong basic groups, at a pH of between 6.9 and 7.4; Stage G: Diafiltration and concentration of the fractions with collagenolytic activity ≧95% originating from stage F, by tangential flow ultrafiltration with 10-50 kD MWCO cassettes; Stage H: Filtration of the solution containing collagenase thus obtained, through an 0.2 μm absolute filter, and storage at a temperature of between −20° and −80° C.; wherein the thus obtained collagenase from