METHOD OF EXTRACTION AND SEPARATION OF SARCOPLASMIC AND MYOFIBRILLAR MEAT PROTEINS TO FRACTIONS BY METHOD OF ONE-DIMENSIONAL ELECTROPHORESIS IN POLYACRYLAMIDE GEL

摘要

FIELD: biotechnology.;SUBSTANCE: method consists in extraction of proteins by homogenisation and centrifugation of the meat samples in buffers-extractants and subsequent fractionation of the resulting extracts in concentrated and separated polyacrylamide gel in the presence of sodium dodecyl sulfate in the electrode buffer. Extraction of sarcoplasmic proteins is carried out using buffer-extractant with pH 7.6, which comprises 0.3 M sucrose, 0.05 M ethylenediaminetetraacetic acid (EDTA), 0.15 M trishydroxymethylaminomethane (Tris). For extraction of myofibrillar proteins the buffer-extractant with pH 7.6 is used, which comprises 0.05 M ethylenediaminetetraacetic acid (EDTA), 0.15 M trishydroxymethylaminomethane (Tris), followed by dialysis of the extract of myofibrillar proteins against solution with pH 7,8, containing 0.6 M potassium chloride (KCl) and 0.1 M trishydroxymethylaminomethane (Tris). After that electro-fractionation of extracts of sarcoplasmic and myofibrillar proteins is carried out in 5% stacking and 11% separating polyacrylamide gel under voltage of 90 V to the stacking gel and 300 V to the separating gel for 2-2.5 hours.;EFFECT: method enables to improve separation of sarcoplasmic and myofibrillar proteins, to reduce the duration of the process of electro- fractionation and enables to separate sarcoplasmic and myofibrillar proteins of pork and beef.;2 dwg, 1 ex