Immobilization method of candida antarctica lipase B

Abstract

The invention discloses an immobilization method of candida antarctica lipase B, and relates to lipase B. The invention provides the immobilization method of the candida antarctica lipase B that lipase is high in storage stability, is facilitated to recycle and is reusable, and is conducive to industrialized application. The immobilization method comprises the steps of 1), firstly performing slant seed culture, activated seed culture and primary seed culture on Pichia pastoris CALB, then performing fermental cultivation by introducing into a fermentation tank containing a BSM culture medium, so as to obtain the candida antarctica lipase B; the preservation number of the Pichia pastoris CALB is CGMCC NO.10277; 2), adding the preprocessed resin into a fermentation tank containing candida antarctica lipase B fermentation liquor, so that the resin is in a suspended state to adsorb; 3) screening out the adsorbed resin in the step 2), washing with water, cooling and drying.

Description

technical field [0001] The invention relates to lipase B, in particular to a method for immobilizing lipase B from Candida antarctica. Background technique [0002] Lipase (EC3.1.1.3) is also called triacylglycerol acyl hydrolase, which can catalyze the decomposition, synthesis, transesterification and transesterification of ester compounds, and can maintain high regio and stereoselectivity. Used in oil hydrolysis, chiral synthesis, food industry and other fields [1] . Candida Antarcticlipase B (CALB) is the most widely used lipase, and its three-dimensional structure and amino acid sequence have been determined by Uppenberg et al. [2] . [0003] Direct application of free lipase in production will bring many problems, such as the existence of free enzyme in a dissolved state, which increases the difficulty of subsequent separation, and the consumption of enzyme increases the production cost. Immobilizing enzymes on carriers is an effective way to maintain their catalyti...

AI Technical Summary

Problems solved by technology

Badgujar [6] immobilized lipase on the biodegradable polymer chitosan, which improved the biocatalytic activity, but after using it for many times, the immobilized enzyme leaked from the carrier and the enzyme activity decreased

Zhang et al. [7] Lipase was successfully immobilized on porous silica particles by covalent cross-linking method, but the carrier of this method needs to undergo tedious chemical treatment, the reaction conditions are harsh, and the cross-linking process leads to partial inactivation of the enzyme

Cao et al. [8] Using sol-gel method to embed and immobilize lipase can effectively improve the enzyme activity, but it is only suitable for the enzyme-catalyzed reaction of substrates and products with relatively small molecular weight

Images