METHOD FOR PREPARING WATER-SOLUBLE IMMOBILIZED ENZYME PROTEASE PREPARATION Russian patent published in 2006 - IPC C12N11/08 A61K38/48 

FIELD: biotechnology, biochemistry, medicine, veterinary science.

SUBSTANCE: method involves preparing a proteolytric enzyme solution being protosubtilin G3H, G10H or G20H mainly in 0.05 M Na-phosphate buffer, pH 7.2-8.2. Prepared solution is purified by saline precipitation of inert proteins by sequential addition of calcium chloride and sodium phosphate followed by keeping the mixture at 8-10°C for 16-18 h and filtration of precipitated deposit through paper and membrane filters. Prepared filtrate is desalted additionally, concentrated and purified by ultrafiltration in hemodialysis columns. The purification process is carried out for two stages by dilution of concentrate with distilled water up to the parent volume and the following repeated concentrating. Polyethylene oxide of molecular mass 1500-4000 Da is added to the prepared highly purified preparation of proteases up to the ratio enzyme - carrier = 1:(3-8). The mixture enzyme - carrier is subjected for repeated sterilizing filtration through membrane filter and irradiated by bremsstrahlung (braking) γ-radiation in the dose 0.5-1.0 Mrad. Invention provides preparing the sterile apyrogenic solution of proteases immobilized on polyethylene oxide with the proteolytic activity 80-120 PU/ml. Invention can be used in preparing injection preparations of immobilized enzymes.

EFFECT: improved preparing method.

4 cl, 1 tbl, 5 ex