METHOD OF PRODUCING HETEROGENEOUS BIOCATALYST BASED ON LIPASE IMMOBILIZED ON CATION-EXCHANGE FIBRES “ВИОН КН-1” IN H-FORM Russian patent published in 2024 - IPC C12N11/00 

FIELD: biotechnology.

SUBSTANCE: disclosed is a method of producing a heterogeneous biocatalyst based on lipase immobilized on ion-exchange fibres. Method comprises adsorption immobilization of lipase in a buffer solution on a cation exchanger matrix during incubation at room temperature with periodic mixing, separating the external solution from the immobilized lipase on the cation exchanger, washing the formed biocatalyst with a buffer; before immobilization, the cation exchanger in the H-form is held for 12 hours at room temperature in a buffer, 20 ml of the buffer is used per 1 g of the cation exchanger, immobilisation is carried out by adding to a suspension of ion-exchange fibres “ВИОН КН-1” in buffer 10 ml of a solution of porcine pancreatic lipase in a buffer in a lipase concentration of 1 mg/ml, buffer solution for immobilisation is 0.05 M phosphate buffer with pH 5.8, followed by incubation with stirring using an electric mixer at rate of 250 rpm for 1.5 hours, then separation of external solution from immobilized lipase on cation exchanger is carried out by centrifugation at 3,000 rpm for 5 minutes and decantation of 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 pore diameter of 25 kDa against 0.2 M 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 amount of 400 ml, and dialysis is continued for another 16 hours until there is no free lipase in the washing solution.

EFFECT: method enables to obtain immobilized lipase with higher hydrolase activity, to expand the number of carriers used for lipase stabilization; besides, the heterogeneous catalyst contains only immobilized lipase and is completely washed from its non-immobilized form.

1 cl, 3 dwg, 1 ex