METHOD OF PRODUCING HETEROGENEOUS BIOCATALYST BASED ON LIPASE IMMOBILIZED ON ANION-EXCHANGE RESINS AV-16-GS AND AN-12P IN OH-FORM Russian patent published in 2024 - IPC C12N11/02 C12N9/00 

FIELD: biotechnology.

SUBSTANCE: invention represents a method of producing a heterogeneous biocatalyst based on lipase immobilized on anion-exchange resins, involving adsorption immobilization of lipase in a buffer solution on an anion exchanger matrix, during incubation at room temperature with periodic mixing, separating the external solution from the immobilized lipase on the anion exchanger, washing the formed biocatalyst with a buffer; before immobilization, the anion exchanger in the OH form is held for 12 hours at room temperature in a buffer, 20 ml of the buffer is used per 1 g of the anion exchanger, immobilisation is carried out by adding to a suspension of anion-exchange resin AV-16-GS or AN-12P 10 ml of a lipase solution from a porcine pancreas in a buffer in a lipase concentration of 3 mg/ml, buffer solution for immobilisation is 0.05 M glycine buffer with pH 10.5, followed by incubation with stirring using an electric mixer at rate of 250 rpm for 1.5 hours, then performing centrifugation at 3,000 rpm for 5 minutes, decantation of the external solution; washing of the formed biocatalyst is carried out by dialysis using a cellophane bag with width of 34 mm, with capacity of 3.7 ml per 1 cm of length, with a pore diameter of 25 kDa against 0.2 M of a phosphate-citrate buffer, pH 6.5, based on the fact that for washing 1 g of obtained biocatalyst 400 ml of 0.2 M phosphate-citrate buffer with pH 6.5 is used, dialysis is carried out for 8 hours, after which the buffer is changed to a portion of buffer in volume of 400 ml and dialysis is continued for another 16 hours until there is no free lipase in the washing solution.

EFFECT: invention enables to obtain a heterogeneous, water-insoluble biocatalyst based on porcine pancreatic lipase with higher activity, due to which it is possible to carry out enzymatic reactions with higher rates.

1 cl, 3 dwg