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Multifunctional enzyme gene HG27 and application

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

Active Publication Date: 2021-06-04
WENS FOODSTUFF GRP CO LTD +1
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  • Summary
  • 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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  • Multifunctional enzyme gene HG27 and application
  • Multifunctional enzyme gene HG27 and application
  • Multifunctional enzyme gene HG27 and 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 experiment

[0042] Feeding Chinese bamboo rats with high-fiber feed such as bamboo, corn cobs, and corn husks for a period of time further induces the efficient proliferation of the cellulose-digesting bacteria in the intestines of bamboo rats. After three weeks of feeding, they were slaughtered, and the microbes of the cecum and colon were collected by the double ligation method on an ultra-clean workbench, and put into liquid nitrogen for quick-freezing and preserved for later use.

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

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

[0045] 2.2. NEB general...

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Abstract

The invention discloses multifunctional enzyme gene HG27 and application. The multifunctional enzyme gene HG27 comprises the following steps of comparing a nucleic acid sequence spliced by rhizomys sinensis intestinal microbe macro transcriptome with a glucoside hydrolase family, screening a sequence with relatively high homology, removing original signal peptide, adding pig parotid gland secretory protein signal peptide, optimizing a pig codon, and connecting to a pcd3.1 vector; transfecting to porcine kidney epithelial cells for expression verification, collecting a cell culture solution for enzyme activity analysis, comparing the enzyme activity, selecting the optimal HG27 gene, and detecting the optimal pH value, protein tolerance and enzymatic reaction of different substrates of the HG27 gene, thereby obtaining the multifunctional enzyme gene HG27 which can be applied to the fields of livestock and poultry feed enzyme preparations, transgenic animal preparation, food and medicine industries and the like, and the problems that cellulose is difficult to degrade and absorb and excrement pollutes the environment are solved.

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 substance 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 the fields of feed, biofuel, fruit juice, etc., where cellulase is a key enzyme used in the biofuel industry. A large number of microorganisms are widely gathered in the intestinal tract of animals, which is considered to be the most effective ecosystem 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 good cellulose-degrading enzymes. Due to the traditional method of simulating natural culture, not only the ...

Claims

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Application Information

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Patent Type & Authority Applications(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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