Animal product-free culture medium for bacteria of the genus clostridium and a process for producing supernatant comprising one or more collagenolytic and gelatinolytic proteases

A process for producing supernatant of Clostridium histolyticum liquid culture comprising one or more proteases with collagenolytic and gelatinolytic activity comprising the following steps:

• a) providing a sterile animal product-free culture medium;
• b) culturing an aliquot of the Clostridium histolyticum stock culture in the culture medium in step (a), under anaerobic conditions at about 37° C. before reaching the stationary growth phase in order to obtain an inoculum;
• c) providing a sterile animal product-free culture medium for Clostridium bacteria according to claim 1;
• d) adding to the culture medium in step (c) a volume of inoculum from step (b) equal to or lower than 10% of the final volume of the medium defined in (c);
• e) culturing the C. histolyticum obtained in (d) under anaerobic conditions at about 37° C. until the stationary growth phase is attained;
• f) removing cellular material and other particulate matter from the liquid phase of the culture in step (e), yielding a supernatant comprising one or more proteases with collagenolytic and gelatinolytic activity.