Method for improving lipase activity persistency

Abstract

The invention provides a method for improving the lipase activity persistency, comprising the steps of: connecting a lipase gene (Genbank No.: AF229435) (with a conventional method) to the N end of a saccharomyces cerevisiae cell wall component FLO sequence (Genbank No.: S73336), then inserting the lipase gene (in a conventional approach) to the C end of a downstream MF alpha1 signal peptide sequence (Genbank No.: M17301) of the GAL1 promoter (Genbank No.: AY428072) of a saccharomyces cerevisiae expression vector so as to construct an expression cassette, which is, from the N end to the C end, as follows: GAL1 promoter + MF alpha1 signal peptide sequence + lipase gene + FLO sequence; shifting a yeast plasmid containing the expression cassette into the saccharomyces cerevisiae cell. According to the invention, the lipase is connected to the saccharomyces cerevisiae cell wall component FLO, so that as long as the saccharomyces cerevisiae cell maintains alive, the lipase can keep active.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for exposing and expressing lipase on the outer surface of the cell wall of Saccharomycescerevisiae to improve the persistence of the enzyme activity. Background technique [0002] Esters are a large class of important products or intermediates widely used in chemical, food, pharmaceutical and other fields. There are hundreds of esters used only as food flavors. The synthesis routes of various esters are the same, and they are prepared by esterification reaction using carboxyl donors (such as acids, acid anhydrides) and hydroxyl donors (such as alcohols, starches, etc.) as raw materials. [0003] Lipases catalyze the biosynthesis of many esters. Compared with the chemical synthesis of esters, the biosynthesis of esters has the advantages of higher efficiency, mild reaction conditions, and low energy consumption. However, the currently used lipase has the problem of insuff...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem of insufficient activity persistence of existing lipase, the present invention provides a technology for making lipase an integral part of Saccharomyces cerevisiae cells, so that lipase can maintain its vitality as long as Saccharomyces cerevisiae cells remain alive, thereby significantly improving the lipase activity. Vitality