METHOD OF PRODUCING A HETEROGENEOUS BIOCATALYST BASED ON LIPASE IMMOBILIZED ON KU-2-8 CATION EXCHANGE RESINS IN THE H-FORM Russian patent published in 2024 - IPC C12N11/02 C12N11/08 C12N9/16 

FIELD: biotechnology.

SUBSTANCE: invention relates to a method of production of a heterogeneous biocatalyst based on lipase immobilized on cation exchange resins, including adsorption immobilization of lipase in a buffer solution onto a cation exchanger matrix during incubation at a room temperature with periodic stirring, separation of the external solution from the immobilized lipase on the cation exchanger, washing the resulting biocatalyst with a buffer; before immobilization, the cation exchanger in the H-form is kept for 12 hours at a room temperature in a buffer, 20 ml of buffer is used per 1 g of cation exchanger, immobilization is carried out by adding 10 ml of a lipase solution to a suspension of the KU-2-8 cation exchange resin in buffer from the pig pancreas in a buffer at a lipase concentration of 1 mg/ml, 0.05 M phosphate buffer with pH 5.8 is used as a buffer solution for immobilization, after which incubation is carried out with stirring using an electric stirrer at a speed of 250 rpm for 1.5 hours, then separation of the external solution from the immobilized lipase on a cation exchanger is carried out by centrifugation at 3,000 rpm for 5 minutes and decantation of the external solution; the resulting biocatalyst is washed by dialysis using a Spectra/Por 6 Standard RC cellophane bag, 34 mm wide, with a capacity of 3.7 ml per 1 cm of length, with a pore diameter of 25 kDa against 0.2 M phosphate-citrate buffer, pH 6.5, based on the calculation that 400 ml of 0.2 M phosphate-citrate buffer with pH 6.5 is used to wash 1 g of the resulting biocatalyst, dialysis is carried out for 8 hours, after which the buffer is changed to a portion of buffer in a volume of 400 ml and dialysis is continued for another for 16 hours until there is no free lipase in the washing solution.

EFFECT: invention makes it possible to obtain a water-insoluble biocatalyst based on lipase from the pig pancreas, immobilized by the adsorption method on a KU-2-8 cation exchange resin in the H-form, including only immobilized lipase and completely washed from its unstabilized (non-immobilized) form, having a higher specific activity, making it possible to carry out enzymatic reactions at higher rates.

1 cl, 3 dwg, 1 ex