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A kind of lipase and its coding gene and application

A lipase gene and coding technology, applied in the field of genetic engineering, can solve problems such as loss of activity and limited application scope.

Active Publication Date: 2016-11-23
SUN YAT SEN UNIV
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  • Claims
  • Application Information

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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  • A kind of lipase and its coding gene and application
  • A kind of lipase and its coding gene and application
  • A kind of lipase and its coding gene and application

Examples

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Embodiment Construction

[0037] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0038] 1: Screening of clones containing lipase sequence

[0039] First compare the deep-sea hydrothermal vent metagenomic sequence with the NCBI non-redundant database to find information similar to the lipase gene; then search for the corresponding sequence information according to the annotation information, design specific primers, and screen the corresponding gene from the library. clones. In order to improve the screening efficiency, we divided the 2880 clones involved in 454 sequencing into 30 groups, that is, each 96-well plate was prepared into a mixed plasmid sample, and another plasmid sample was prepared from a mixture of 2880 clones as a positive for screening As a control, in this way, the first round of screening is performed using the 30 mixed samples as PCR amplification templates, and the number of the mixed sample c...

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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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/55C12N15/11C12N15/70C12N1/21C12N9/20C12R1/19
Inventor 徐安龙王磊任政华吴珊陈尚武邹琼付永贵
Owner SUN YAT SEN UNIV
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