METHOD FOR PRODUCING proteinase of positively charged protein fraction in a growing population ESCHERICHIA COLI

摘要

A method of producing proteinases positively charged protein fractions in the growing population of Escherichia coli, comprising the preservation of cells in the presence of 80-90% glycerol buffered with subsequent removal of the cell walls 3% Triton X-100 extraction increasing salt concentrations: 0.14 M, 0.35 M; 2 M NaCl, 6 M guanidine hydrochloride with 0,1% β-mercaptoethanol, release of the above fractions of positively charged proteins by ion exchange chromatography with Amberlite IRC-50 in a discontinuous gradient of guanidine hydrochloride 6%, 8.9%, 10.6 % 13% 0,1 M potassium phosphate buffer pH 6.8, elution of proteinases by the method of affinity chromatography on a column of Sepharose 4 B with immobilized trypsin inhibitor followed by evaluation of proteolytic activity.