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Plant lactobacillus plantarum tannase gene and application thereof

A plant lactobacillus, tannase technology, applied in the direction of plant gene improvement, application, genetic engineering, etc., can solve the problems of low purity of tannase, low catalytic activity of ester bonds and depsi acid bonds, etc., to improve catalytic activity. , the effect of high catalytic activity and simple operation

Active Publication Date: 2020-07-10
四川省危险化学品质量监督检验所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is that the existing tannase has low purity and low catalytic activity to ester bonds and depsipphenolic acid bonds. Tannase with high yield and high catalytic activity towards ester bond and depside bond

Method used

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Experimental program
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Effect test

Embodiment 1

[0025] The acquisition of the Lactobacillus plantarum tannase gene Tg and its mutant gene MTg: PCR was performed with the Lactobacillus bacterium liquid as a template, and the full-length sequence of the Lactobacillus plantarum tannase gene Tg was cloned (as shown in SEQ ID NO. 1 in the sequence table). (Shown); using the cloned full-length sequence of the Lactobacillus plantarum tannase gene Tg, site-directed mutation was performed using PCR technology to obtain the full-length sequence of the mutant gene MTg (as shown in SEQ ID NO. 2 in the sequence table).

Embodiment 2

[0027] Application of Lactobacillus plantarum tannase gene Tg and its mutant gene MTg in tannase production:

[0028] 1) Application of Lactobacillus plantarum tannase gene Tg in the preparation of tannase:

[0029] The Lactobacillus plantarum tannase gene Tg was quickly expressed in E. coli by the method of gene recombination, and the expressed tannase was purified by affinity chromatography and ion exchange chromatography to obtain high yield and high purity. Lactobacillus plantarum tannase; Take 1 liter of LB liquid medium as an example, the purified tannase yield> 80mg, purity> 95%, the determination of the activity of the tanninase showed that the catalysis activity of the Lactobacillus plantarum tanninase on the ester bond was 204 U / mg, and the activity on the depsip bond was 25 U / mg.

[0030] 2) Application of Lactobacillus plantarum tannase mutant gene MTg in the preparation of tannase:

[0031] 1. Construction of recombinant expression plasmid

[0032] Synthesize the modified ...

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Abstract

The invention discloses a plant lactobacillus plantarum tannase gene and application thereof. The nucleotide sequence of the plant lactobacillus plantarum tannase gene Tg is shown in SEQ ID NO.1, thenucleotide sequence of a plant lactobacillus plantarum tannase mutant gene MTg is shown in SEQ ID NO.2, and the mutant gene MTg is obtained by transforming gene fragments of the plant lactobacillus plantarum tannase gene Tg by using a fixed point mutation technology. By adopting the plant lactobacillus plantarum tannase gene and the mutant gene thereof, oriented production and application of tannase with a high yield and high purity can be achieved, and a mutant gene expression product has high catalysis activity upon ester bonds and depside bonds.

Description

Technical field [0001] The invention relates to a gene and its application, in particular to the Lactobacillus plantarum tannase gene and its application. Background technique [0002] Tanninase, also known as tanninyl hydrolase, belongs to serine hydrolase. It has esterase and depsilase activities. It can degrade the depsid and ester bonds in tannins, release gallic acid and glucose, and make tannins Enzymes are widely used in food, animal feed, brewing, chemical and pharmaceutical industries. [0003] Different tanninases have different catalytic activities on ester bonds and depsilon bonds. The traditional method of producing tanninase is mainly through liquid submerged fermentation and solid-state fermentation. The production of tanninase by these two methods takes longer Long, high cost, not conducive to purification, and the produced tannase has an unclear catalytic activity on ester bonds and depsil bonds, and the catalytic efficiency is low, resulting in great limitations ...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N9/18C12N15/70
CPCC12N9/18C12N15/70C12Y301/0102
Inventor 王晓云官青松暨磊王姣丽陈伟
Owner 四川省危险化学品质量监督检验所
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