METHOD FOR PREPARING COLLAGENASE PREPARATION Russian patent published in 2005 - IPC

FIELD: biotechnology, preparative biochemistry, enzymes.

SUBSTANCE: invention relates to a method for preparing the highly purified and highly active enzyme preparation collagenase. Method for preparing the collagenase preparation involves homogenization of hepatopancreas tissue of industrial species crabs in buffer aqueous solutions, isolation of collagenase solution, its purification by successive two-stage ultrafiltration on membranes providing cutting off inert substances and lyophilic drying of the preparation. Ultrafiltration is carried out on poly-fibrous membranes with size of pores 15 kDa and concentrate prepared at the 1-st stage of ultrafiltration is diluted with water in the ratio = 1:(5-10), and concentrate prepared at the 2-d stage of ultrafiltration is subjected for additional successive microfiltration on membranes with size of pore 100 kDa and ultrafiltration on membranes with size of pores 15 kDa. Homogenization of hepatopancreas of industrial species crabs is carried out in acetate-ammonium buffer containing calcium acetate at pH 6.4-6.7 followed by precipitation of collagenolytic enzymes with ammonium sulfate at 60-70% degree of saturation. Invention provides increasing the total collagenolytic activity, specific activity of the preparation and its purity. The collagenase preparation can be used in medicine, veterinary science, biochemistry and food industry and for research aims also.

EFFECT: improved preparing method.

2 cl, 2 ex