FIELD : biotechnology.SUBSTANCE : invention relates to biotechnology and medicine, in particular to industrial technology for producing Collalysinenzyme preparation for medical use. Method of producing Collalysinby culturing producer Clostridium histolyticum in anaerobic conditions on Ramon nutrient medium with subsequent separation of microbial mass by filtration, wherein for cultivation producer is used Clostridium histolyticum 468 or producer Clostridium histolyticum SK B-8362, and separation of microbial mass is carried out by means of cascade of depth filters with pore diameter from 3.0–0.8 to 0.3–0.1 mcm with exclusion of culture in native solution, using nominal and absolute sterilizing filtration of native solution with further ultrafiltration, concentration and chromatographic purification based on Sephacryl S-100 sorbent. Use of Collalisinfor treating fibroproliferative diseases.EFFECT : method for preparing said preparation enables to obtain a preparation having high collagenase activity and preserving stability of all quality indices during the specified shelf life of 2 years.6 cl, 1 dwg, 1 ex