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Bacillus licheniformis for producing 2,3,5,6-tetramethylpyrazine in high yield as well as separation culture method and application of bacillus licheniformis

A technology of Bacillus licheniformis and tetramethylpyrazine, which is applied in the field of high-yield Bacillus licheniformis and its isolation and cultivation, can solve the problem of energy consumption, long period of koji making process, and difficult tetramethylpyrazine production. Problems such as high-efficiency accumulation of azine, to achieve the effect of improving production efficiency and far-reaching social and economic significance

Active Publication Date: 2020-06-02
JIANGSU KINGS LUCK BREWERY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the process of koji making and accumulation is an open fermentation mode, and the composition and structure of microorganisms are relatively complex, so it is difficult to control the process parameters to regulate the efficient accumulation of tetramethylpyrazine
At the same time, the koji making process has a long cycle (30 days), low efficiency, and mainly an acidic environment, which is not conducive to the efficient synthesis of tetramethylpyrazine, resulting in a relatively low tetramethylpyrazine content
At the same time, the pure culture of bacterial bran koji can increase the content of tetramethylpyrazine, but the bran koji needs to be cooked in this process, which consumes a certain amount of energy

Method used

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  • Bacillus licheniformis for producing 2,3,5,6-tetramethylpyrazine in high yield as well as separation culture method and application of bacillus licheniformis
  • Bacillus licheniformis for producing 2,3,5,6-tetramethylpyrazine in high yield as well as separation culture method and application of bacillus licheniformis
  • Bacillus licheniformis for producing 2,3,5,6-tetramethylpyrazine in high yield as well as separation culture method and application of bacillus licheniformis

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Effect test

Embodiment 1

[0065] The present embodiment provides a kind of Bacillus licheniformis high-yielding tetramethylpyrazine, and the separation method comprises the following steps:

[0066] (1) Mix high-temperature daqu and sterile saline to obtain a bacterial suspension;

[0067] (2) Dilute the bacterial suspension obtained in step (1) and treat it at 80°C for 20 minutes and then cultivate it on beef extract peptone solid medium to obtain colonies; beef extract peptone solid medium includes beef extract 3g / L, peptone 10g / L, sodium chloride 5g / L, agar powder 20g / L, solvent is water; pH=7.2;

[0068] (3) pick the morphologically regular bacterium colonies in step (2) and carry out streak culture on the beef extract peptone solid medium to obtain a single pure-bred bacterium colony;

[0069] (4) Pick a single colony for liquid culture, prepare a seed solution, inoculate it on a bran solid medium and cultivate it at 37°C for 24 hours, measure the content of tetramethylpyrazine, and obtain potent...

Embodiment 2

[0071] The present embodiment provides a kind of Bacillus licheniformis high-yielding tetramethylpyrazine, and the separation method comprises the following steps:

[0072] (1) Mix high-temperature daqu and sterile saline to obtain a bacterial suspension;

[0073] (2) Dilute the bacterial suspension obtained in step (1) and treat it at 85°C for 15 minutes and then cultivate it on beef extract peptone solid medium to obtain colonies; beef extract peptone solid medium includes beef extract 2g / L, peptone 12g / L, sodium chloride 6g / L, agar powder 22g / L, solvent is water; pH=7.3;

[0074] (3) pick the morphologically regular bacterium colonies in step (2) and carry out streak culture on the beef extract peptone solid medium to obtain a single pure-bred bacterium colony;

[0075] (4) Pick a single colony for liquid culture, prepare a seed solution, inoculate it on a bran solid medium and cultivate it at 37°C for 24 hours, measure the content of tetramethylpyrazine, and obtain potent...

Embodiment 3

[0077] The Bacillus licheniformis that embodiment 1 screens obtains carries out the observation of bacterium colony form and individual form, the result is as follows figure 1 and figure 2 as shown ( figure 1 It is the colony form figure of the bacillus licheniformis of high-yielding tetramethylpyrazine of the present invention, figure 2 It is the individual morphology diagram of Bacillus licheniformis with high tetramethylpyrazine production in the present invention). Depend on figure 1 It can be seen that the colonies of Bacillus licheniformis with high tetramethylpyrazine production in the present invention are flat, with irregular edges, white, and rough and wrinkled surfaces. Depend on figure 2 It can be seen that: under an optical microscope, the Bacillus licheniformis individual shape of the high-yielding tetramethylpyrazine of the present invention is rod-shaped, has a single existence, and also has paired and chain-like existences, and these above-mentioned cha...

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Abstract

The invention relates to bacillus licheniformis for producing 2,3,5,6-tetramethylpyrazine in a high yield as well as a separation culture method and application of the bacillus licheniformis. The bacillus licheniformis for producing the 2,3,5,6-tetramethylpyrazine in the high yield is named a bacillus licheniformis S1001 strain, the preservation unit is China General Microbiological Culture Collection Center, the preservation time is February 25, 2019, the preservation number is CGMCC No.17255, and the address is No.3, Yard NO.1 Beichen West Road, Chaoyang District, Beijing. The strain has thecharacteristics of producing the 2,3,5,6-tetramethylpyrazine in the high yield by using a raw material under the condition of facultative anaerobic solid fermentation, and when the strain is appliedto a raw material solid culture process, starter rich in the 2,3,5,6-tetramethylpyrazine can be produced, so that the strain has an important effect on further improving healthy active substances in awine body.

Description

technical field [0001] The invention belongs to the technical field of microbial fermentation and relates to a bacillus licheniformis and its isolation and cultivation method and application, in particular to a high-yielding tetramethylpyrazine bacillus licheniformis and its isolation and cultivation method and application. Background technique [0002] Tetramethylpyrazine (2,3,5,6-tetramethylpyrazine), also known as Ligustrazine, is the first alkaloid extracted from Ligusticum Chuanxiong, a commonly used traditional Chinese medicine, in my country. It has been proven to protect the liver, dilate blood vessels, improve microcirculation and inhibit platelet aggregation, etc. It is widely used in the treatment of cardiovascular and cerebrovascular diseases, glomerular diseases and respiratory diseases. A certain concentration of tetramethylpyrazine is also detected in Chinese liquor, which is mainly produced through the joint action of microorganisms and Maillard heat reaction ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12G3/021C12R1/10
CPCC12N1/20C12N1/02C12G3/021C12R2001/10C12N1/205
Inventor 唐群勇吴建峰季方祝赛峰胡烨烨
Owner JIANGSU KINGS LUCK BREWERY
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