METHOD FOR PRODUCTION OF THIOL:PROTEIN-DISULFIDE OXIDOREDUCTASE FROM PLANT RAW MATERIALS

摘要

FIELD: enzymology.;SUBSTANCE: claimed method includes production of homogenate from wheat Triticum aestiivum L. coarse grinding flour by extraction of protein structure with aqueous solution containing EDTA and DTT/ Extraction is carried out at continuous agitation of flour and aqueous solution in ratio of 1:3. Obtained extract is separated from solid phase by centrifugation at 3000-5000 rmp for 15-30 min/ Obtained supernatant is twice fractionated. The first fractionation is carried out at 20 % saturation of solution with ammonium sulfate; the second one is carried out at 70 % saturation followed by deposit separation using centrifugation after the first and second centrifugation. Obtained deposit id is resuspended in 0.1 M tris-HCl buffer containing EDTA and separated by centrifugation at 10000-14000 rpm. Isolated target crude product is salted out on column with Sefadex. Extract is purified by ion-exchange chromatography on column with Sefadex DEAE A50 in 0.1 M tris-HCl buffer followed by rechromatography of target product on column with Sefadex DEAE A50 in 0.1 M tris-HCl buffer, containing EDTA. Purified extract is freeze-dried.;EFFECT: enzyme with various purification ratio.;11 cl, 2 dwg, 1 tbl, 2 ex