Acid-resistant and high temperature-resistant beta-mannase gene and application thereof

A technology of mannanase and high temperature resistance, applied in the field of enzyme engineering

Inactive Publication Date: 2012-08-08
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the use of genetic engineering to industrialize the production of aci

Method used

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  • Acid-resistant and high temperature-resistant beta-mannase gene and application thereof
  • Acid-resistant and high temperature-resistant beta-mannase gene and application thereof
  • Acid-resistant and high temperature-resistant beta-mannase gene and application thereof

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Embodiment Construction

[0098] The present invention is described in further detail below in conjunction with embodiment.

[0099] This embodiment includes the following steps:

[0100] (1) Extraction of total DNA of Bacillus subtilis CGMCC NO1.1628

[0101] 1) Pick a single colony of Bacillus subtilis CGMCC NO1.1628 strain and inoculate it into LB liquid medium, and incubate at 37°C for 12 hours; 2) Pipette 1.4 mL of the bacterial solution into a centrifuge tube, centrifuge at 10,000 rpm for 30 seconds, discard the medium, and repeat Repeat several times until the bacterial solution is completely collected; 3) Resuspend the bacteria in 480 μL STE solution (0.1 mol / L NaCl, 10 mmol / L Tris-HCl, 1 mmol / L EDTA), and place at room temperature for 30 min; 4) Add 20 μL Lysozyme and RNase A, placed at 37°C for 1h; 5) Add 1 / 10 volume of 10% SDS, 0.1 mg / mL proteinase K (final concentration) and place at 55°C for 2h; 6) Add 1 / 3 volume after cooling to room temperature 5mol / L NaCl and 1 volume of chloroform, p...

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Abstract

The invention discloses an acid-resistant and high temperature-resistant beta-mannase gene and application thereof. The beta-mannase gene has the nucleotide sequence shown as SEQ ID No. 1 as well as the amino acid sequence shown as SEQ ID No. 2. The invention also discloses the application of the acid-resistant and high temperature-resistant beta-mannase gene to the preparation of an acid-resistant and high temperature-resistant beta-mannase. The optimum pH value and the optimum temperature of the beta-mannase are 4.6 and 70 DEG C respectively; the beta-mannase has certain characteristics of acid resistance and high temperature resistance, and is particularly applied to industries, such as foods, feeds, paper making, printing and dyeing, and textile.

Description

technical field [0001] The invention belongs to the field of enzyme engineering, and in particular relates to an acid-resistant and high-temperature resistant β-mannanase gene and its application in preparing acid-resistant and high-temperature resistant β-mannanase. Background technique [0002] β-mannanase (β-1,4-mannan mannohydrolase, EC 3.2.1.78), also known as β-D-mannanase or β-1,4-D-mannanase, is a class of Endohydrolase capable of hydrolyzing mannan oligosaccharides and mannan polysaccharides (including mannan, galactomannan, glucomannan, etc.) containing β-1,4-D-mannosidic bonds[ Zhimin He, Jun Zhang , Dongpo Huang. A kinetic correlation for konjac powder hydrolysis by β-mannanase from Bacillus licheniformis[J]. Biotechnology Letters, 2001, 23: 389-393. ]. It belongs to hemicellulase, a broad-spectrum inducible multifunctional enzyme with cellulase activity, widely present in animals, plants and microorganisms[ Vendula ValáValá?ková,ková, Petr Baldrian. Estimatio...

Claims

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Application Information

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IPC IPC(8): C12N15/56C12N15/63C12N1/21C12N9/42C12R1/19
Inventor 毛绍名章怀云
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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