Cold adapted endo beta-xylanase gene XynA and use

A xylanase and gene technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of less research on polar sea ice microorganisms, difficult sampling and cultivation, etc.

Inactive Publication Date: 2009-07-15
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the difficulties of sampling and culturing, there are relatively few studies on polar sea ice microorganisms.

Method used

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  • Cold adapted endo beta-xylanase gene XynA and use
  • Cold adapted endo beta-xylanase gene XynA and use
  • Cold adapted endo beta-xylanase gene XynA and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] The gene fragment sequence encoding the cold-adapted endoxylanase XynA is shown as SEQ ID NO.1. The corresponding relationship between nucleotides and encoded amino acids in genes is as described above. The genome DNA fragment has a total of 1272bp, which contains a 1272bp open reading frame encoding cold-adaptive endoxylanase XynA, the start codon is located at 1bp, and the stop codon is located at 1270bp, encoding a total of 423 amino acids.

Embodiment 2

[0106] Embodiment 2: gene fragment cloning method of the present invention

[0107] Source of strain: Glaciecola mesophila DSM 15026T is isolated from marine invertebrates. Deposited in the German Culture Collection DSMZ, accession number: DSM 15026T. bought in the market.

[0108] The specific method is as follows:

[0109] 1. Cloning of gene encoding cold-adapted endoxylanase XynA

[0110] 1.1 Extraction of Genomic DNA from Glaciecola mesophila SM0901

[0111] Refer to "Refined Molecular Biology Experiment Guide".

[0112] (1) Cultivate 50ml of bacterial culture to saturation, take 15ml of culture and centrifuge at 12000g for 5min;

[0113] (2) Add 5670 μl TE to the precipitate, and resuspend it by repeatedly blowing with a pipette. Add 300μl 10% SDS and 30μl 20mg / mL proteinase K, mix well, and incubate at 37°C for 2.5-3h;

[0114] (3) Add 1000μl 5M NaCl, mix thoroughly, then add 800μl CTAB / NaCl solution, mix well, and incubate at 65°C for 30-60min (from this step, po...

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Abstract

The invention relates to a gene XynA of cold adapted endo-beta-xylanase and an application thereof. The genome DNA fragment sequence is shown as SEQ ID N o. 1. The cold adapted beta-xylanase gene XynA is applicable for preparing beta-xylanase XynA. The obtained beta-xylanase XynA is used as neutral cold adapted hydrolase for hydrolyzing oat xylan, beech xylan and birch xylan to produce xylobiose and xylotriose. Optimum enzyme activity temperature is 30 DEG C and optimum pH is 7.0.

Description

technical field [0001] The invention relates to a gene of a novel cold-adaptive endo-xylanase, which belongs to the field of biotechnology. Background technique [0002] β-xylanase is a glycoside hydrolase secreted by bacteria and fungi that can degrade xylan. Most of them belong to the 10th and 11th families. In recent years, a few β-xylanases have been found to belong to the 8th family. . The properties of the β-xylanases of the 10th family from different sources are not much different; however, the enzymes of the 11th family have strong substrate specificity, low molecular weight, and the properties of the enzymes of different branches are quite different. The β-xylanases of family 10 can hydrolyze glycosidic bonds adjacent to branch points and non-reducing end glycosidic bonds. For β-xylanases of family 10, two consecutive non-substituted xylose residues between two branch points are sufficient for their action, while enzymes of family 11 require 3 such residues. base...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/56C12N9/24C12P19/12C12P19/00C12R1/01
Inventor 张玉忠陈波郭兵陈秀兰俞勇周百成
Owner SHANDONG UNIV
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