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Bacillus subtilis for efficiently producing beta glucanase as well as application thereof

A technology of Bacillus subtilis and glucanase, applied in the field of bioengineering, can solve the problems of poor stability and low enzyme activity, and achieve the effects of high activity, stable activity and good genetic stability

Active Publication Date: 2014-01-29
杭州皇冠农业生物工程技术研究中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a bacillus subtilis with high production of β-glucanase, which solves the problems of low enzyme activity and poor stability of traditional enzyme-producing strains

Method used

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  • Bacillus subtilis for efficiently producing beta glucanase as well as application thereof
  • Bacillus subtilis for efficiently producing beta glucanase as well as application thereof
  • Bacillus subtilis for efficiently producing beta glucanase as well as application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 Obtaining of high-yield bacterial strains

[0061] 1. LiCl-ultraviolet compound mutagenesis

[0062] The starting strain was Bacillus subtilis QB-01, which was preserved by the School of Life Sciences of Zhejiang University and presented by Shao Jianzhong (Zhejiang, Hangzhou).

[0063] After the starting strain was activated, it was prepared with sterile physiological saline to a concentration of 10 6 cells / mL single-cell suspension; take 10mL single-cell suspension in a culture dish with a magnetic rotor, add 600 μL of 10% LiCl solution, and mix well; open the lid of the culture dish, and use a 15W ultraviolet lamp to Irradiate for 75s; under dark conditions, dilute the mutagenized single cell suspension by 10 3 times, spread it on a screening medium plate, and culture it in the dark at 37°C for 48 hours in a constant temperature incubator to obtain a LiCl-ultraviolet compound mutagenic strain.

[0064] Using the unmutated bacterial suspension as a cont...

Embodiment 2

[0108] Fermentation culture and enzyme activity assay of embodiment 2 bacterial strains

[0109] The Bacillus subtilis CGMCC 6897 selected in Example 1 was inoculated in the liquid seed medium at 37°C, 220r / min, shaken for 24h, and the seed culture solution was inserted into the shake flask with an inoculum size of 10% (V / V). In the fermentation medium of 37 DEG C, 220r / min, culture 48h, measure β-glucanase enzyme activity, β-glucanase enzyme activity assay method is the same as embodiment 1. Modify the corresponding factors according to the specific experimental conditions below. (1) Optimization of fermentation medium formula

[0110] 1. Carbon source optimization

[0111] Carbon source is the main component of fermentation medium. Can provide energy for microbial growth and metabolism. Different microorganisms have different physiological characteristics and have great differences in their ability to utilize various carbon sources.

[0112] The basal medium 1 supplemen...

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Abstract

The invention discloses bacillus subtilis for efficiently producing beta glucanase as well as an application thereof. The bacillus subtilis is named as Bacillus subtilis pj101. The strain has been already stored in China General Microbiological Culture Collection Center on January, 28, 2012, wherein the preserving number is CGMCC No.6897. The invention further provides an application of the bacillus subtilis strain for efficiently producing beta glucanase in fermenting production of beta glucanase. The bacillus subtilis strain CMGCC 6897 is good in genetic stability, and beta glucanase produced is high and stable in activity. In addition, the fermenting condition of the strain is further optimized to obtain a better enzyme producing effect, so that bacillus subtilis can be applied to industrial fermenting production of beta glucanase.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a strain of bacillus subtilis with high production of β-glucanase and its application. Background technique [0002] β-glucanase is a general term for a class of hydrolytic enzymes that can degrade β-glucan in grains, mainly including 4 enzymes, namely endo-β-1,3-glucanase, endo-β-1 ,3-1,4-glucanase, exo-β-1,3-glucanase and exo-β-1,4-glucanase. As a specific exogenous enzyme, β-glucanase is widely used in beer brewing industry and feed industry. β-glucanase can decompose β-glucan gel in barley and malt, reduce the viscosity of wort, increase the filtration rate and yield of wort, help to improve the flavor of beer and maintain the non-biological stability of finished wine; Adding it to grain feed can reduce the viscosity of the animal's digestive tract, effectively improve the digestion and absorption of nutrients by monogastric animals, increase the feed conversion rate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/24C12R1/125
Inventor 胡向东朱静许美芳叶茂
Owner 杭州皇冠农业生物工程技术研究中心有限公司
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