Beta-glucanase as well as coding gene IDSGH5-26 and application thereof

A glucanase and gene technology, applied in the field of genetic engineering, can solve problems such as insufficient acid resistance of β-glucanase, achieve good industrial application value, good stability, and ensure novelty effects

Active Publication Date: 2021-01-08
ZHEJIANG WANLI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently obtained β-glucanase still has defects,

Method used

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  • Beta-glucanase as well as coding gene IDSGH5-26 and application thereof
  • Beta-glucanase as well as coding gene IDSGH5-26 and application thereof
  • Beta-glucanase as well as coding gene IDSGH5-26 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] 1. Extraction of total RNA from rumen contents of Hu sheep

[0040] (1) Rapidly grind the rumen content samples frozen at -80°C into powder in liquid nitrogen, and add 1ml of RNApure Ultra-Pure Total RNA Quick Extraction Kit (Adline Biotechnology Co., Ltd., Beijing) for every 50-100mg sample , China) in the lysate RL and then homogenate. The sample volume cannot exceed 10% of the RL volume;

[0041] (2) Vigorously shake and mix the homogenate sample on a vortex shaker to lyse the cells, and incubate at 15-30°C for 5 minutes to completely decompose the ribosomes;

[0042] (3) Centrifuge at 12,000 rpm for 10 minutes at 4°C to remove insoluble matter, and carefully transfer the supernatant to a new RNase-free centrifuge tube;

[0043] (4) Add 0.2ml chloroform to each ml of lysate RL, shake vigorously for 15s, centrifuge at 12000rpm, 4°C for 10min;

[0044] (5) Transfer the upper colorless aqueous phase to a new tube, add absolute ethanol half the volume of the aqueous p...

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Abstract

The invention relates to glucanase, in particular to beta-glucanase as well as a coding gene IDSGH5-26 and application thereof, and belongs to the field of gene engineering. The nucleotide sequence ofthe beta-glucanase gene IDSGH5-26 for coding the beta-glucanase derived from rumen microorganisms of Hu sheep is as shown in SEQ ID No. 1. The optimum temperature of the beta-glucanase is 50 DEG C, and the beta-glucanase keeps good stability under the condition of 40 DEG C. The optimum pH value of the enzyme is 5.0, the enzyme has high pH tolerance, 74% or above of activity can be kept when the pH value is 4.00-10.0, and products obtained after beta-glucan is degraded by the enzyme are monosaccharide and oligomerization cellooligosaccharide with the polymerization degree being 2-4. The recombinant beta-glucanase has a good industrial application value.

Description

technical field [0001] The field of the invention relates to a glucanase, in particular to a β-glucanase, its encoding gene IDSGH5-26 and its application, belonging to the field of genetic engineering. Background technique [0002] β-glucan is a linear polysaccharide formed by hundreds or even thousands of β-D-glucose residues linked by β-1,3 and β-1,4-glucosidic bonds, and is the main structure of the cell wall of grasses It is one of the components of sexual polysaccharides, accounting for about 5.5% of its dry matter, and it is also widely present in the cell walls of bacteria, yeasts, macrofungi and other microorganisms. As a kind of non-starch polysaccharide (NSP), β-glucan is an important anti-nutritional factor in plant feed. It has high viscosity and is not easy to be degraded, and it is difficult to be digested and absorbed by monogastric animals such as pigs and poultry, which will lead to a decrease in the utilization rate of grains and related feeds, and greatly...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/56C12N15/70C12N15/11C12P19/14C12P19/12C12P19/02
CPCC12N9/2434C12N15/70C12P19/14C12P19/12C12P19/02
Inventor 王谦王佳堃高德英曹佳雯何波
Owner ZHEJIANG WANLI UNIV
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