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Alkaline xylanase as well as encoding gene and application thereof

A technology of xylanase and alkaline, which is applied in the field of alkaline xylanase and its coding gene and application, and can solve problems such as the inability to produce heat-resistant alkaline xylanase on a large scale

Inactive Publication Date: 2015-05-20
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The development of industrial application of xylanase has been carried out internationally, but only a small number of companies can produce xylanase for the paper industry, and there is still room for improvement in its alkali resistance and heat resistance; Systematic research on heat-resistant alkaline xylanase, but it is also impossible to produce heat-resistant alkaline xylanase on a large scale

Method used

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  • Alkaline xylanase as well as encoding gene and application thereof
  • Alkaline xylanase as well as encoding gene and application thereof
  • Alkaline xylanase as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] (1) Materials

[0059] 1. Strains

[0060] Alkaliphilic Micrococcus is an alkaline xylanase-producing strain Micrococcus sp.S1 screened from Shenzhen mangrove soil. The strain preservation unit: China Center for Type Culture Collection (CCTCC), preservation date: June 27, 2014, preservation address: China. Wuhan. Wuhan University, preservation number: CCTCC NO: M 2014295. E.coli TOP10F' was purchased from Invitrogen;

[0061] 2. Carrier

[0062] The Escherichia coli cloning vector pMD18-T was purchased from Dalian Bao Biological Company, and the Escherichia coli expression vector pET-28a(+) (Novagen, KanR) was purchased from Novagen Company; figure 1 , its multiple cloning site map is shown in figure 2 .

[0063] 3. Medium

[0064] (1) Selection medium: xylan 8.0g / L, KNO 3 1.0g / L, MgSO 4 ·7H 2 O 0.5g / L, NaCl 15g / L, KH 2 PO 4 1.5g / L, solid medium plus agar 15-20g / L, pH 9.0;

[0065] (2) LB medium: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, solid medi...

Embodiment 2

[0156] (1) Materials

[0157] 1. Strains

[0158] Alkaliphilic Micrococcus sp.S1 strain (CCTCC No: M 2014295). The host bacteria E.coli BL21Star(DE3) and E.coli TOP10F' were purchased from Invitrogen;

[0159] 2. Carrier

[0160] Escherichia coli expression vector pET-28a(+) (Novagen, Kan R ) was purchased from Novagen; vector map see Figure 9 , its multiple cloning site map is shown in Figure 10 .

[0161] 3. Medium and buffer

[0162] (1) Selection medium: xylan 8.0g / L, KNO 3 1.0g / L, MgSO 4 ·7H 2 O 0.5g / L, NaCl 15g / L, KH 2 PO 4 1.5g / L, solid medium plus agar 15-20g / L, pH 9.0;

[0163] (2) LB medium: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, solid medium plus agar 15-20g / L, pH 9.0, autoclaved at 121°C for 20min.

[0164] (3) TE buffer: 10mmol / L Tris-Hcl, pH8.0, 1mmol / L EDTA, pH8.0.

[0165] (4) Alkaline lysis solution I, II, III (plasmid extraction): Alkaline lysis solution I: glucose 50mmol / L, Tris-HCl (pH 8.0) 25mmol / L, EDTA 10mmol / L. About 100mL pe...

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PUM

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Abstract

The invention discloses alkaline xylanase as well as an encoding gene and application thereof. The S1 gene sequence is a complete open reading frame (ORF), the open reading frame starts with an initiation codon ATG and ends up with a termination codon TAA and comprises 609 nucleotides in all, and the nucleotide sequence of the open reading frame is as shown in SEQ ID No:1. The nucleotide sequence of the protein of the gene code of alkaline xylanase S1 is as shown in SEQ ID No:2. The invention further discloses application of the alkaline xylanase S1 in industries such as paper making.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an alkaline xylanase, its coding gene and application. Background technique [0002] Hemicellulose is the second largest renewable resource on earth after cellulose. Xylan is a hemicellulose linked by β-1,4 linkages with the unit of xylanose, which is abundant in broad-leaved trees and most annual plants. The content of hemicellulose in nature accounts for 30% of biomass, second only to cellulose. [0003] Xylan is a heteropolysaccharide whose main chain is composed of xylopyranose linked by β-1,4 glycosidic bonds. The so-called xylanase is endo-β-1,4-D-xylanase (endo-β-1,4-D-xylanase) [EC3.2.1.8], its main function is random It cleaves the β-1,4-glycosidic bond between the xylopyranose residues in the backbone of xylan to generate xylose, xylooligosaccharides or xylooligosaccharides with side chains of different lengths. Because xylanase plays an important role in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N15/56C12N15/70C12N1/21D21C9/10
CPCC12N9/2482C12Y302/01008D21C9/10
Inventor 邢苗刘森林陈伟钊
Owner SHENZHEN UNIV
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