METHOD FOR PRODUCING OF YEAST ALCOHOL OXIDASE Russian patent published in 1995 - IPC

FIELD: biotechnology. SUBSTANCE: method involves cultivation of yeast as producer of alcohol oxidase (AO). Collected cells of cultivated biomass are suspended. Thus prepared suspension is disintegrated, cellular homogenate is centrifuged, supernatant liquid which contains AO and catalase being purified. Said purification is carried out by sorption of cellular-free centrifugate, 0.01-0.04 M tric-HCl buffer together with 0.05 M NaCl, 0.1 mM EDTA and 0.1 mM dithioteitole being used. Said process takes place at pH 7.6-8.2, it is followed by eluation of protein with the help of 0.1-0.5 M NaCl, mentioned above buffer being used. The latter process is carried out at pH7.6-8.2 in the presence of 0.1 mM EDTA and 0.01 mM dithioteitole. Fractions by 4 ml are collected at rate 8 ml/h. Thus ferment preparation of AO having no catalase impurities is prepared. Desired product yield is 43.4 % , its specific activity is 12.5 units. EFFECT: improves efficiency of the method. 6 dwg, 3 tbl