METHOD OF PREPARING CHOLESTEROL-ESTERASE, TRIPSIN, DEOXYRIBONUCLEASE, AND RIBONUCLEASE FROM BOVINE PANCREAS

摘要

FIELD: pharmaceutical chemistry and biotechnology.;SUBSTANCE: invention aims at biotechnological preparation of following medicaments: pancreatic ribonuclease, pancreatic deoxyribonuclease, tripsin, and cholesterolesterase. Method envisages grinding bovine pancreas and homogenation thereof. Resulting homogenate is extracted with cooled ethanol at constant stirring, centrifuged, and subjected to second extraction, now with sulfuric acid containing magnesium sulfate, at constant stirring. Extract containing desired enzymes is separated from precipitate via centrifugation. Ballast proteins are precipitated by ammonium sulfate at 30-35% saturation and removed from supernatant via centrifugation. Cholesterolesterase is precipitated by ammonium sulfate at 45-50% saturation of extract, tripsin at 60-65% saturation, and DNA-ase at 95-98% saturation followed by dialysis and chromatographic purification on DEAE-cellulose column.;EFFECT: enabled preparation of four enzymes in a single technological process, increased yield of pancreatic ribonuclease and increased its specific activity.;6 dwg, 8 tbl