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A kind of bacillus licheniformis beta-mannanase mutant and its application

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

Active Publication Date: 2022-05-13
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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  • A kind of bacillus licheniformis beta-mannanase mutant and its application
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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

The present invention selects Bacillus licheniformis β-mannanase gene, mutates the gene through error-prone PCR, and performs high-efficiency secretory expression in Bacillus subtilis, and screens to obtain a mutant with improved enzyme activity and pH stability. The results showed that the optimum temperature of β-mannanase and its mutants was 60°C, and the optimum pH was 6.0. After 4 days of fermentation, the mutant enzyme activity reached 13888±260 U / mL, which was 2.2 times that of the wild type. The half-life was 30 minutes. In addition, the β-mannanase mutant had higher pH stability than the wild type, and the residual enzyme activity was higher than 80% after incubation at 37°C for 1 hour in the range of pH 5.0-10.0. The enzyme has shown a good application prospect in aquaculture.

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 ...

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Application Information

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Patent Type & Authority Patents(China)
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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