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A kind of multifunctional enzyme gene hg27 and its application

A multi-functional enzyme, HG27 technology, applied in the field of genetic engineering, can solve the problems of polluted environment, difficult to degrade and absorb cellulose, etc., achieve high cellulase activity, strong trypsin tolerance, and improve the effect of gastrointestinal environment

Active Publication Date: 2022-07-22
WENS FOODSTUFF GRP CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, a cellulase and glucanase bifunctional enzyme gene HG27 was obtained, which can be used in livestock and poultry feed enzyme preparations, transgenic animal preparation, food and pharmaceutical industries, etc., to solve the problem of cellulose refractory absorption and excretion environmental pollution problem

Method used

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  • A kind of multifunctional enzyme gene hg27 and its application
  • A kind of multifunctional enzyme gene hg27 and its application
  • A kind of multifunctional enzyme gene hg27 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] The invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0041] 1. Animal feeding test

[0042] Feeding Chinese bamboo rats with high-fiber feeds such as bamboo, corn core and corn peel for a period of time further induces the efficient proliferation of the cellulose-digesting flora in the intestinal tract of bamboo rats. After three weeks of rearing, they were slaughtered, and the cecum and colonic microorganisms were collected by double ligation method on the ultra-clean workbench, and put into liquid nitrogen for quick-freezing storage for future use.

[0043] 2. RNA extraction and metatranscriptome sequencing and sequence analysis of bamboo rat gut microbes

[0044] 2.1. Use QIAGE's ZR Soil / Fecal RNA MicroPrep (R2040) to extract intestinal microbial RNA from the cecum and colon of Chinese bamboo rat, and the electrophoresis results are as follows figure 1 shown.

[0045] 2.2. NEB general library cons...

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Abstract

By comparing the spliced ​​nucleic acid sequence of the macrotranscriptome of the intestinal microorganisms of Chinese bamboo rat with the glycoside hydrolase family, the invention selects the sequence with higher homology, removes its original signal peptide and adds the pig parotid gland secretory protein signal peptide, It was linked to the pcd3.1 vector after porcine codon optimization. It was transfected into pig kidney epithelial cells for expression verification, the cell culture medium was collected for enzyme activity analysis, the enzyme activity was compared, the optimal HG27 gene was selected, and the optimal pH, protein tolerance and enzymatic reaction of different substrates were detected. . Thus, a multifunctional enzyme gene HG27 is obtained, which can be applied in the fields of livestock and poultry feed enzyme preparations, preparation of transgenic animals, food and pharmaceutical industries, etc., to solve the problems of refractory absorption of cellulose and environmental pollution by excreta.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a multifunctional enzyme gene HG27 and its application. Background technique [0002] Cellulase is the main constituent of lignocellulose, the most abundant renewable resource, and is ubiquitous in plant cell walls rich in hemicellulose and lignin. However, due to its insolubility and crystal structure, it is difficult to be widely used in feed, biofuel, fruit juice and other fields, among which cellulase is the key enzyme used in the biofuel industry. A large number of microorganisms are widely accumulated in the intestinal tract of animals, which are considered to be the most effective ecosystems for degrading cellulose, such as herbivorous animals such as cattle, pigs, and bamboo rats. Therefore, looking for cellulase from animal gut microbes may be one of the best ways to find excellent cellulose-degrading enzymes. Due to the traditional method of simulating natural cultu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/56C12N15/62C12N9/42C12N9/46C07K19/00C12N15/85C12N5/10C12N1/21A23L33/17A23L33/135A23K20/189A23K10/18A61K48/00A61K38/47A61P1/00
CPCC12N9/2437C12N9/2454C12N15/85C12N5/0686C12Y302/01011A23L33/17A23L33/135A23K20/189A23K10/18A61K48/005A61K38/47A61P1/00C07K2319/02C12N2800/107C12N2800/22C12N2510/02A23V2002/00A23V2200/30A23V2200/32
Inventor 吴珍芳张献伟黄广燕王豪强杨善欣莫健新蔡更元
Owner WENS FOODSTUFF GRP CO LTD
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