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Bacillus tequilensis and application thereof

A technology of bacillus tequila and bacillus, applied in the field of bioengineering, can solve problems such as restricting the development of beer and feed industries

Active Publication Date: 2012-10-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the enzyme preparations used in China are domestically produced except for a small amount of feed enzyme preparations. Most of the enzyme preparations rely on imported products. Since the production technology is a commercial secret, this greatly limits the development of the beer and feed industry. Industrial enzyme preparation technology with independent intellectual property rights is of great significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1 Bacillus tequila screening and enzyme activity assay method

[0014] Take a certain sample from the fragments of wasp nests and add it to 30mL sterile saline, break up the glass beads to make a suspension, and treat at 80°C for 10min. The suspension was serially diluted, spread on an MRS plate supplemented with 2% Congo red, and incubated in a 37°C incubator for 24h. Pick a single colony conspicuous by the Congo red discoloration circle. Streak separation on the plate was repeated three times to obtain a purified single colony for identification in the next step.

[0015] Identification is mainly through: 1) Observation of colony characteristics: directly observe with the naked eye, and select bacterial colonies with a diameter of 1-1.5mm, round shape, smooth surface with slightly wrinkled, milky white, etc. that meet the characteristics of Bacillus colonies. 2) Observation of individual morphology: Observation by Gram staining with an optical microscope....

Embodiment 2

[0017] Example 2 Process for producing β-1,3-1,4-glucanase by fermentation

[0018] The culture medium used for producing β-1,3-1,4-glucanase by Bacillus tequila fermentation is divided into seed medium and fermentation medium. Wherein the seed culture medium is LB culture medium: Tryptone 10g L -1 , Yeast Extract 5g L -1 , NaCl 10g L -1 , pH7.0. The main components of the fermentation medium are: corn flour 40g L -1 , bean cake powder 61.1g L -1 , barley flour 68.4g L -1 , dipotassium hydrogen phosphate 1g L -1 , calcium chloride 0.1g L -1 , magnesium sulfate 1g L -1 , high temperature amylase 16μL / L. The filling volume is 30mL per 250mL Erlenmeyer flask. The medium sterilization condition is 121°C, 20min.

[0019] Pick a single colony of Bacillus tequila from the plate and activate it on the slant for 8 hours, pick a ring of bacteria from the slant and inoculate it into LB medium, and culture it in a shaker flask at 37°C and 200rpm for 15 hours to the middle and l...

Embodiment 3

[0020] Example 3 Applied to the Construction of Genetically Engineered Bacteria

[0021] According to the bglS design in NCBI in Bacillus amyloliquefaciens LL3 (NC_017190.1), primers were cloned and the β-1,3-1,4-glucanase gene was used to construct an expression plasmid, which can be expressed in Escherichia coli or Pichia pastoris A series for efficient fermentative expression and production of β-1,3-1,4-glucanase.

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PUM

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Abstract

The invention discloses bacillus tequilensis CGX5-1 which is preserved with the number CGMCC No.5935 in China General Microbiological Culture Collection Center on March 26, 2012. The bacillus tequilensis has high capability of producing beta-1, 3-1, 4-glucanase, and the bacillus tequilensis and good genes thereof are applied to the field of production of industrial enzyme preparations and have good prospects in reduction of cost for producing the beta-1, 3-1, 4-glucanase and improvement on enzyme preparation quality.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a strain of Bacillus tequila and its application. technical background [0002] There are many kinds of Bacillus, which are widely distributed, and mainly exist in soil, water, air and the intestinal tract of animals. And extracellular enzymes are abundant, which has a good research prospect. Due to the complexity of the growth environment, Bacillus will also secrete and produce cellulase, such as β-glucanase. β-glucanase is widely used in beer brewing: using β-glucanase in the saccharification process can reduce the viscosity of wort, increase the filtration speed, and reduce energy consumption; in addition, in wort Adding β-glucanase can also reduce the glucan content and prevent beer turbidity, thereby improving the abiotic stability of beer. Due to the high content of β-glucan in wheat feed, it hinders the digestion and absorption of active ingredients in the feed in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/21C12N9/24C12R1/07
Inventor 王金晶刘晓玲李崎李永仙刘春凤郑飞云
Owner JIANGNAN UNIV
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