Lipase, and coding gene and application thereof

Abstract

The invention relates to a lipase gene, a lipase protein coded by the lipase gene, and a latent application of the protein in the fields of industrial production and medicine synthesis due to the oil hydrolysis property of the protein. A clone corresponding to the lipase is screened from a metagenome library with note information of Fosmid library gene information fragments obtained after 454 sequencing of a deep sea hydrothermal derived sample Fosmid library as a research material, and the Fosmid clone is sequenced through a shotgun combined walking method to obtain lipase coding ORF with the gene sequence represented by a <1> sequence in a sequence table. The lipase target gene is inserted into a vector pET-32a(+) expression vector, and expression and purification are carried out to obtain the mature peptide of the gene coding polypeptide. The amino sequence of the peptide is represented by a <2> sequence in the sequence table. Enzyme activity researches of the lipase show that the lipase has good lipase characteristics and has latent application values in the fields of industrial production and medicine synthesis.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a lipase, its encoding gene and its application. Background technique [0002] Lipase (E.C.3.1.1.3), also known as triacylglycerol hydrolase, is a special ester bond hydrolase that can catalyze the hydrolysis of natural substrate oil to generate fatty acids, glycerol, and monoglyceride or diester. It widely exists in In prokaryotes (such as bacteria) and eukaryotes (such as molds, mammals, plants, etc.). Studies have shown that in addition to catalyzing the hydrolysis and synthesis of glyceride compounds, it can also be used to catalyze transesterification reactions, biosurfactant synthesis, peptide synthesis, polymer synthesis, and drug synthesis. The stereospecificity of lipase can also catalyze the separation of optical isomers and the synthesis of chiral drugs which are difficult to complete by chemical methods. Therefore, lipase has been widely used in many fields such a...

AI Technical Summary

Problems solved by technology

[0003] As a biocatalyst, the surrounding environment (temperature, pH, organic reagents, etc.) has a great influence on the activity of the enzyme, and many enzymes will lose their activity under some harsh reaction conditions (high temperature, low temperature or strong acid-base environment). This limits its scope of application

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