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A kind of alkaline xylanase and its coding gene and application
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A xylanase, alkaline technology, applied in the field of alkaline xylanase and its encoding gene and application, can solve the problem of inability to large-scale industrial production of heat-resistant alkaline xylanase and the like
Inactive Publication Date: 2017-10-03
SHENZHEN UNIV
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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
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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.coliTOP10F' 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;
[0158] Alkaliphilic Micrococcus sp.S1 strain (CCTCC No: M 2014295). The host bacteria E.coliBL21Star(DE3) and E.coli TOP10F' were purchased from Invitrogen;
[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 (pH8.0) 25mmol / L, EDTA 10mmol / L. About 100mL per ...
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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...
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