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Bacillus licheniformis beta-mannase mutant and application thereof

A technology of Bacillus licheniformis and mannanase, which is applied in the direction of application, glycosylase, enzyme, etc., can solve the problems of difficult market demand and low yield of Bacillus licheniformis, and achieve the effect of efficient secretion and expression

Active Publication Date: 2021-02-26
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

Derived from Bacillus licheniformis β-mannanase, it has good thermal stability and alkali stability, and has potential application value in feed, food, papermaking and other industries, but the production of this enzyme in Bacillus licheniformis is low, and it is difficult to meet the market need

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  • Bacillus licheniformis beta-mannase mutant and application thereof
  • Bacillus licheniformis beta-mannase mutant and application thereof
  • Bacillus licheniformis beta-mannase mutant and application thereof

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Embodiment Construction

[0025] The technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0026] 1 Materials and methods

[0027] 1.1 Materials and reagents

[0028] The plasmids and bacterial strains used in the present invention are shown in Table 1 and Table 2 respectively. Fastpfu, Hifi Taq, easyTaq, dNTP, etc. are products of Beijing Quanshijin Biotechnology Co., Ltd., and other materials are domestic reagents. Biometra Tone PCR instrument is a product of Jena Company, Germany, SpectraMaxm i3x multifunctional microplate reader.

[0029] Table 1 Plasmids used in the present invention

[0030] .

[0031] Remarks: [1] Wu SC and Wong SL., Development of improved pUB110-based vectors for expression and secretion studies in Bacillus subtilis Journal of biotechnology 1999, 72:185-195.

[0032] Table 2 Bacterial strains used in the present invention

[0033] .

[0...

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Abstract

According to the invention ,the bacillus licheniformis beta mannase gene is selected, the gene is mutated through error-prone PCR, efficient secretory expression is carried out in bacillus subtilis, and a mutant with improved enzymatic activity and pH stability is obtained through screening. Results show that the optimum temperature of beta mannase and the mutant thereof is 60 DEG C, the optimum pH value is 6.0, the enzyme activity of the mutant reaches 13888 + / -260 U / mL when the mutant is fermented for 4 days and is 2.2 times that of a wild type, the half-life period at 70 DEG C is 30 min, inaddition, the beta mannase mutant has higher pH stability than the wild type, and after incubating at 37 DEG C for 1h in the range of pH5.0-10.0, the residual enzyme activity is higher than 80%. Theenzyme shows a good application prospect in the breeding industry.

Description

technical field [0001] The invention relates to a bacillus licheniformis beta-mannanase mutant and its application. Background technique [0002] β-mannanase (EC 3.2.1.78) can randomly hydrolyze the β-1,4-mannosidic bond of mannan to produce low molecular weight mannan oligosaccharides. It is widely used in feed, food, medicine, papermaking, textile and other industries It has wide application value. β-mannanase can reduce the anti-nutritional factor effect of mannan and improve the utilization value of livestock feed; on the other hand, its degradation product mannan oligosaccharide can promote the growth of lactic acid bacteria and bifidobacteria, which is beneficial to the intestinal health of animals , It also has the functions of enhancing animal immunity, regulating glucose and lipid metabolism, promoting growth and antioxidation. [0003] β-mannanase widely exists in plants, animals and microorganisms, but the yield of enzymes from natural sources is low, and it is ...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12N15/56C12N15/75C12N1/21C12R1/125
CPCC12N9/2494C12N15/75C12Y302/01078
Inventor 张春晓高伟强陈晓波王丽丽
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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