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Improved xylanase with high specific activity, its gene expression vector and recombinant yeast cell, and expression method

A technology of xylanase and expression vector, applied in the field of genetic engineering, can solve problems such as general heat resistance, and achieve the effect of good heat stability

Active Publication Date: 2009-05-27
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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Problems solved by technology

But its heat resistance is generally

Method used

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  • Improved xylanase with high specific activity, its gene expression vector and recombinant yeast cell, and expression method
  • Improved xylanase with high specific activity, its gene expression vector and recombinant yeast cell, and expression method
  • Improved xylanase with high specific activity, its gene expression vector and recombinant yeast cell, and expression method

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Experimental program
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Embodiment

[0039] Experimental conditions

[0040] 1. Strains, vectors and genes Escherichia coli strain E.coli DH 5a , plasmid pET-22b(+), etc. were purchased from Promega Company, yeast strain Pichia pastoris GS115 (His - Mut + ), plasmid pPIC9 is a product of Invitrogen Company. Xylanase gene xynB (EMBL accession number is: AJ292317) is obtained from Streptomycesolivaceoviridis (preservation number: CGMCC No.1348) by this laboratory, depository unit: General Microbiology Center of China Committee for Culture Collection of Microorganisms, referred to as: CGMCC, depository unit address: It was cloned in No. 13, North Yiyi Road, Zhongguancun, Haidian District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, 100080, date of preservation: April 11, 2005).

[0041]2. Enzymes and kits Restriction enzymes, ligases, Taq enzymes, and DNAaseI are products of Boehringer. PCR Kits were purchased from Promega.

[0042] 3. Biochemical reagents DNA synthesis reagents are product...

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Abstract

The present invention provides one kind of improved xylanase with high specific activity and its coding gene and efficient expression method. The improved xylanase has raised heat stability, and after treating at 80 deg.c for 1 min, the improved xylanase XYNB has residual enzyme activity over 98 %. The present invention also provides recombinant yeast cell containing the xylanase gene of the present invention, and the recombinant yeast has xylanase expressing amount up to 10000 IU / mL fermentation liquid. The xylanase of the present invention may be used widely in feed and food industry.

Description

technical field [0001] The invention relates to the field of genetic engineering. Specifically, the present invention relates to a molecularly improved xylanase and its gene. The present invention also relates to an expression vector containing the xylanase gene, a recombinant yeast cell, and a high-efficiency expression method of the gene. Background technique [0002] Xylan (xylan) is a hybrid five-carbon sugar, the main chain is connected by multiple xylopyranosyl groups through xylosidic bonds. A variety of short substituents of different sizes are attached to the side chains. Xylan mainly exists in the secondary wall of plant cells and acts as a link between lignin and other polysaccharides. Xylan is an important component of plant hemicellulose, which accounts for one-third of the total plant carbohydrates, and is the second most abundant renewable material resource after cellulose in nature. The amount of xylan contained in different plants is also different. Gener...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/56C12N9/24C12N9/42C12N15/63C12N15/81C12N15/80C12N15/82C12N15/85C12N1/19
Inventor 姚斌罗会颖王亚茹袁铁铮史秀云柏映国
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI