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Alkaline xylanase, and encoding gene and application thereof

A xylanase and alkaline technology, applied in the field of alkaline xylanase and its coding gene and application, can solve the problem that there are not many excellent strains with high enzyme activity

Inactive Publication Date: 2020-06-09
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although there is a certain foundation for the research on xylanase at home and abroad, there are still not many excellent strains with high enzyme activity that can be produced on a large scale so far.

Method used

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  • Alkaline xylanase, and encoding gene and application thereof
  • Alkaline xylanase, and encoding gene and application thereof
  • Alkaline xylanase, and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Materials

[0047] (1) Bacterial species: Bacillus alkalophilus, which is a strain Bacillus sp.WMN1 with alkaline xylanase activity obtained from soil screening. The strain deposit unit: China Center for Type Culture Collection (CCTCC), deposit date: January 3, 2020, deposit address: China. Wuhan. Wuhan University, deposit number: CCTCC NO: M 2020007. The host strain E.coli DH5α was purchased from Invitrogen.

[0048] (2) Carrier:

[0049] Escherichia coli cloning vector pMD19-T Vector with Amp r Resistance markers were purchased from Takara (Dalian) Bao Biological Company.

[0050] (3) culture medium

[0051] LB medium: tryptone 10g, yeast extract 5g, NaCl 10g, add 15-20g agar to the solid medium, dilute to 1L, autoclave at 121°C for 20min.

[0052] 1% agarose gel: Dissolve 1g agarose in 100mL 1×TAE buffer, then add 5μL Goldview.

[0053] Amp: Dissolve 1g of ampicillin sodium salt in a sufficient amount of water, and finally dilute to 10mL. After filter ster...

Embodiment 2

[0160] (1) Materials

[0161] 1. Bacterial species: Bacillus sp. WMN1 (CCTCC NO: M 2020007). The host strain E.coli DH5α and the expression host strain E.coli BL21 Star (DE3) were purchased from Invitrogen.

[0162] 2. Vector: Escherichia coli cloning vector pMD19-T Vector, with Amp r Resistance sign. Escherichia coli expression vector pET-28a(+) contains Kan r Resistance markers were purchased from Takara (Dalian) Bao Biological Company.

[0163] 3. Medium and buffer

[0164] LB medium: tryptone 10g, yeast extract 5g, NaCl 10g, add 15-20g agar to the solid medium, dilute to 1L, autoclave at 121°C for 20min.

[0165] 1% agarose gel: Dissolve 1g agarose in 100mL 1×TAE buffer, then add 5μL Goldview.

[0166] 100mM IPTG: 2.4g IPTG dissolved in 100mLddH 2 In O, 0.45μm membrane filter was sterilized, aliquoted, and stored at -20°C.

[0167] Amp: Dissolve 1g of ampicillin sodium salt in a sufficient amount of water, and finally dilute to 10mL. After filter sterilization, al...

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Abstract

The invention discloses alkaline xylanase, and an encoding gene and an application thereof, belonging to the technical field of biology. The sequence of WMN1 gene of the alkaline xylanase is a complete open reading frame (ORF) which starts with an initiation codon ATG and ends with a termination codon TAA and comprises 1308 nucleotides in total; and the nucleotide sequence of the open reading frame is shown as SEQ ID No: 6. The amino acid sequence of the protein encoded by the WMN1 gene of the alkaline xylanase is shown as SEQ ID No: 7. The invention also discloses an application of the WMN1 gene of the alkaline xylanase in industries of papermaking, washing and the like.

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] Foreign research on xylanase began in the 1950s, and it is relatively complete from strain construction to product development and production, and industrialized mass production of xylanase can be used. The current research hotspots mainly focus on the cloning and expression of microbial xylanase genes, the regulation mechanism and induction, purification and identification of xylanase, and the preparation of immobilized enzymes by nanotechnology. The research on xylanase in our country is relatively late, and the current research is mainly on the screening and domestication of excellent xylanase-producing strains, the optimization of enzyme production conditions, the purification of enzymes and the study of physical and chemical properties, xylanase gene cloning, expression and recombin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N15/56C12N15/70C11D3/386
CPCC11D3/38636C12N9/2482C12N15/70C12Y302/01008
Inventor 刘森林钟雅芳陈伟钊
Owner SHENZHEN UNIV