METHOD OF PRODUCING MULTI-ENZYME PREPARATION Russian patent published in 2011 - IPC C12N9/60 C12N9/04 

FIELD: chemistry.

SUBSTANCE: method involves culturing yeast in a bioreactor on a molasses medium with addition of milk whey in amount of 50% of the total volume of the culture medium. Yeast cells are separated from the culture fluid by centrifuging and subjected to transformation on a ball mill at temperature 25±2°C at rotor speed 10000 rpm for 2.0 ±0.3 hours. The transformed yeast cells are extracted with 0.06 mM phosphate buffer with pH 8.5 and temperature 37±2°C, followed by centrifuging to obtain supernatant fluid. The obtained supernatant fluid is treated twice with cooled acetone solution to obtain a precipitate. The obtained precipitate is dissolved in distilled water and subjected to dialysis against distilled water for 3 hours at temperature +4±2°C and centrifuged to obtain supernatant fluid. The obtained supernatant undergoes microfiltration on membranes with pore size 0.22 mcm and ballast proteins are removed via ion-exchange chromatography on a column with CM and DEAE cellulose.

EFFECT: invention enables to obtain an ethanol-oxidising multi-enzyme preparation with high degree of purity and high catalytic activity and increase output of yeast biomass.

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