Novel multifunctional enzyme gene HG32 and application

A multifunctional enzyme and gene technology, applied in the field of genetic engineering, can solve the problems of refractory cellulose degradation and absorption, excrement pollution of the environment, etc.

Active Publication Date: 2021-05-28
WENS FOOD GRP CO LTD +1
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

Thus, a cellulase gene HG32 was obtained, which can be used in livestock and poultry feed enzyme preparations, transgenic animal preparation

Method used

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  • Novel multifunctional enzyme gene HG32 and application
  • Novel multifunctional enzyme gene HG32 and application
  • Novel multifunctional enzyme gene HG32 and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0043] The invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0044] 1. Animal feeding experiment

[0045] Feeding Chinese bamboo rats with high-fiber feed such as bamboo, corn husks, and corncobs for a period of time further induces the efficient proliferation of bacteria that digest cellulose 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.

[0046] 2. RNA extraction, metatranscriptome sequencing and sequence analysis of gut microbes in bamboo rats

[0047] 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.

[0048] 2.2. NEB general li...

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Abstract

The invention discloses a novel multifunctional enzyme gene HG32 and application, the enzyme is derived from Chinese rhizomyidae enteric microorganisms, can be compatible with an eukaryotic cell expression system, expresses cellulase with high biological activity, and can be applied to the fields of livestock and poultry feed enzyme preparations, transgenic animal preparation, food and medicine industries and the like; the invention solves the problems that plant cellulose is difficult to degrade, the feed nutrition utilization rate is low and the like are solved, and pollution of nitrogen, organic matter and the like in animal excrement. According to the invention, the application comprises the following steps: sequencing a bamboo rat intestinal microbe macro transcriptome, and splicing the sequences; performing homology comparison with the existing glycoside hydrolase family on NCBI, and screening high-expression suspected cellulase genes with homology higher than 50%; predicting the signal peptide, and replacing the original signal peptide with the pig parotid gland secreted protein signal peptide; optimizing according to pig codon preference, and cloning to an eukaryotic expression vector; transfecting into pig kidney epithelial cells through lipidosome for expression; collecting a cell culture supernatant; and carrying out enzyme activity analysis and comparison.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a novel multifunctional enzyme gene HG32 and its application. Background technique [0002] Cellulose is an abundant green carbon source in nature and the main component of plant cell walls. An appropriate amount of cellulose is beneficial to animal intestinal peristalsis and promotes feces excretion. However, cellulose is also the main component of plant cell walls. Most livestock and poultry are monogastric animals such as pigs and chickens. Their stomachs and small intestines lack cellulase, which makes it difficult to destroy plant cell walls and hinder the full absorption of intracellular nutrients. These Undigested nutrients enter the natural environment with excrement, causing environmental pollution. Naturally, cellulose often exists in a supramolecular aggregate structure and is difficult to be degraded. The way to solve this problem is to find excellent cellulose de...

Claims

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Application Information

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IPC IPC(8): C12N15/56C12N9/42C12N15/85A23K20/189A23K50/75A01K67/027
CPCC12N9/2437C12N15/8509A23K20/189A23K50/75A01K67/0278C12N2800/22A01K2207/15A01K2217/072A01K2227/108A01K2267/02
Inventor 张献伟黄广燕王豪强莫健新杨善欣吴珍芳蔡更元
Owner WENS FOOD GRP CO LTD
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