Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus

A technology of tetramethylpyrazine yield and bacillus, applied in the field of bioengineering, can solve the problems of genetic stability and safety hazards, difficult industrial production, high production cost, etc., and achieve low cost, simple operation, safe and stable yield Effect

Inactive Publication Date: 2016-08-10
QILU UNIV OF TECH +1
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AI Technical Summary

Problems solved by technology

Since the synthesis mechanism of TTMP in brewing is still unclear, the productivity of fermenting TTMP by wild strains using its own metabolic pathway is low; adding precursor substances (3-hydroxybutanone) can effectively strengthen the metabolic pathway, but the production cost is high , low cost-effectiveness; through the optimization of fermentation medium and culture conditions and the regulation of its fermentation process, the yield of TTMP in the fermentation system can be effectively increased, but there are constraints such as product inhibition; genetic modification of the strain can also be effective Increase the yield of TTMP, but the molecularly modified strains are used in the production of food fermentation, there are hidden dangers of genetic stability and safety, and it is difficult to carry out industrial production

Method used

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  • Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus
  • Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus
  • Method for improving output of tetramethylpyrazine produced through fermentation by Bacillus

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Experimental program
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Effect test

Embodiment 1

[0022] Embodiment one: application of pH control fermentation improves the ability of bacillus to produce TTMP

[0023] Step 1: Preparation of seed solution: Under sterile conditions, use an inoculation loop to inoculate a loop of activated Bacillus species into a 250 mL Erlenmeyer flask containing 50 mL LB medium, shake at 37 °C and 200 r / min Bed shaking culture for 12 h, then transferred to a 250 mL Erlenmeyer flask containing 50 mL of LB medium containing 10 g / L glucose according to the inoculum size 2 % (V / V), shaken on a shaker at 37 °C and 200 r / min After culturing for 12 h, adjust the cell concentration (OD 600 ) is 0.05, as the seed solution.

[0024] Step 2: Inoculate the seed liquid into a 500 mL Erlenmeyer flask containing 200 ml of YPG medium containing 70 g / L glucose at an inoculum volume (V / V) of 4%, and culture it with shaking at 37°C and 200 r / min. Add 5ml of glucose solution with a concentration of 1mg / ml at 12h. From the 72h, add 10mol / L NaOH to control the...

Embodiment 2

[0031] Embodiment 2: The ability of adding acetaldehyde to regulate TTMP production in the initial stage of application fermentation

[0032] According to the preparation of seed liquid in Example 1 and the fermentation method when controlling the pH value of the fermented liquid to be 7.5, acetaldehyde of gradient concentration was added to the medium at the initial stage of fermentation: 1.0, 2.0, 4.0 and 8.0 g / L, and the fermentation test was carried out to When TTMP reaches its peak value, samples are taken at regular intervals during the fermentation process to analyze the concentration of bacteria in the fermentation broth (OD 600 ), TTMP and 3-hydroxybutanone content.

[0033] 1) As the amount of acetaldehyde increased from 0 g / L to 1.0 g / L, Bacillus licheniformis ( B. lincheniformisBL1) The highest yield of TTMP increased from 37.89 g / L to 43.13 g / L. When the addition of acetaldehyde in the medium was increased above 1.0 g / L, the yield of the fermentation product TTM...

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Abstract

The invention specifically relates to a method for improving output of tetramethylpyrazine produced through fermentation by Bacillus, belonging to the field of bioengineering. The method is characterized in that a certain amount of acetaldehyde is added into a fermentation medium in an early stage of fermentation and NaOH is added to allow the pH value of fermentation broth to be slightly greater than 7.0, thereby improving the output of tetramethylpyrazine by Bacillus. The method has the advantages that the output of tetramethylpyrazine is increased by adding acetaldehyde in the early stage of fermentation; the output of tetramethylpyrazine is substantially increased through controlling of a weakly alkaline condition; and the method is economic, environment-friendly and low in cost.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for improving the yield of tetramethylpyrazine fermented by bacillus. Background technique [0002] Tetramethylpyrazine (TTMP), also known as Ligustrazine, is an effective active ingredient in the traditional Chinese medicine Ligusticum Chuanxiong, which can lower blood pressure and has significant curative effects in the treatment of cerebrovascular, pulmonary edema, and kidney diseases, and is widely used in pharmaceutical synthesis , has been widely used clinically as a standing drug. TTMP is another important aroma substance, which naturally exists in liquor, cocoa beans, tea, beef, mutton, pork, cheese, and nuts. spices. At present, the research on pyrazine compounds in the liquor industry has paid more attention. It has been preliminarily confirmed that in addition to the Maillard pathway of TTMP in liquor, microbial metabolite pathways can also produce ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12R1/07C12R1/10C12R1/125
CPCC12P17/12
Inventor 孟武王瑞明王松江王腾飞王婷
Owner QILU UNIV OF TECH
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