Acid-resistant bacillus velezensis DQA21 for producing acetoin and application

A technology of DQA21 and Bacillus, applied in the field of bioengineering, can solve the problems of unmentioned difference, strain ACT yield needs to be further improved, unmentioned strain growth, aroma production, etc., to achieve high ACT yield, fast fermentation speed, increase The effect of flavor substances

Active Publication Date: 2021-01-22
WULIANGYE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned prior art does not mention whether the strain is still suitable for growth and aroma production under the acidic conditions of the brewing process, and neither mentions the difference between the flavor produced by the strain and the liquid fermentation under the condition of solid-state fermentation
In addition, the ACT production of the strain needs to be further improved

Method used

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  • Acid-resistant bacillus velezensis DQA21 for producing acetoin and application
  • Acid-resistant bacillus velezensis DQA21 for producing acetoin and application
  • Acid-resistant bacillus velezensis DQA21 for producing acetoin and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Isolation and screening of Bacillus velezensis DQA21

[0044] 1) Sample processing

[0045] Weigh 1 g of Daqu, put it into a Erlenmeyer flask filled with 99 mL of sterile water and contain sterile glass beads, oscillate to disperse Daqu evenly, and prepare a sample suspension. The suspension was heated in a water bath at 85°C for 10 minutes to eliminate non-spore-forming bacteria, and then 1 mL of the suspension was added to the enrichment medium for overnight enrichment culture.

[0046] Liquid enrichment medium in g / L: glucose 30g / L, peptone 10g / L, yeast powder 5g / L, sodium chloride 0.5g / L, wine fermented grain extract, pH 6.0, sterilized at 121°C for 20min . Among them, put 10g of fermented grains in a 250mL triangular flask, add 50mL of water, stir well, and soak at room temperature (25°C) for 15min; during the soaking time, stir once every 5min; Make up to 100mL.

[0047] Two) separation and purification

[0048] Use a pipette gun to draw 1mL of the...

Embodiment 2

[0055] Example 2: 16S rDNA identification and physiological and biochemical experiments

[0056] 1) Identification: First, through physiological and biochemical experiments, it was preliminarily judged to be Gram-positive short bacillus, and then identified by 16SrDNA molecular biology technology to confirm that the strain was Bacillus velezensis and named it Bacillus velezensisDQA21.

[0057] 2) ACT standard curve drawing

[0058] Concentrations of 0.05, 0.1, 0.2, 0.4, 0.8g / L ACT standard solutions were prepared respectively. Pipette 0.1mL of ACT standard solutions of different concentrations into test tubes, add 4.5mL of creatine mixture, shake and mix well, react in a water bath at 30°C for 30min, and measure the OD value at 522nm. The ACT standard curve was drawn with the ACT concentration as the abscissa and the absorbance value as the ordinate. Such as figure 1 As shown, its regression equation is y=5.3989x-0.0444, and the correlation coefficient R 2 =0.9991, indicat...

Embodiment 3

[0070] Example 3: Optimization of fermentation conditions for high-yield ACT strains

[0071] 1) Single factor test

[0072] The glucose concentrations were selected to be 40, 60, 80, 100, 120g / L, the pHs were 5.0, 5.5, 6.0, 6.5, 7, the temperatures were 31, 34, 37, 40, 43°C, and the inoculum sizes were 3, 5, 7, 9, 11%, the speeds are 120, 150, 180, 210, 240r / min, the filling volume is 30, 40, 50, 60, 70mL / 250mL triangular flask and other 6 factors to do single factor test , 3 parallel experiments for each group.

[0073] Depend on Image 6 a It can be seen that with the increase of glucose concentration in the fermentation medium, the yield of ACT fermented by Bacillus velezensis DQA21 increases significantly. When the glucose concentration reached 80g / L, the ACT output reached 18.372g / L. When the glucose concentration was greater than 100g / L, the ACT production obviously declined, and it can be seen that the higher sugar concentration has an inhibitory effect on the grow...

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Abstract

The invention discloses acid-resistant bacillus velezensis DQA21 for producing acetoin and application. The acid-resistant bacillus velezensis DQA21 was preserved in the China Center for Type CultureCollection (CCTCC) with the preservation number being CCTCC NO:M 2020360 on July 27, 2020. The fermentation conditions of the strain are optimized, and the optimal fermentation conditions are as follows: the glucose concentration is 80 g / L, the pH is 5.46, the temperature is 37.31 DEG C, the liquid loading amount is 49.17 mL, the inoculation amount is 5%, and the rotating speed is 210 r / min. Underthe conditions, the average yield of the acetoin fermented by a strain shake flask is 25.60 g / L, and is improved by 39.36% compared with the yield of 18.37 g / L of the strain before optimization. Thestrain has the advantages of high acetoin yield, high fermentation speed and the like, can be used for producing baijiu, and can be used for increasing the content of the acetoin in the baijiu, improving the mouthfeel and flavor and enhancing the quality of the baijiu.

Description

technical field [0001] The invention relates to the technical field of bioengineering, more specifically, it relates to an acid-resistant acetoin-producing (Acetoin, ACT) bacillus DQA21 (Bacillus velezensis DQA21) and its application. Background technique [0002] Qu is the bone of wine, and the quality of distiller's koji directly affects the quality, output and style of liquor. Medium-high temperature Daqu is a saccharification starter for brewing Luzhou-flavor liquor. It contains various microorganisms such as yeast, mold, and bacteria. style characteristics. The highest product temperature of medium-high temperature Daqu can reach 62°C during the process of making koji. Because Bacillus has the characteristic of using spores to cope with the stress environment, Bacillus has become one of the dominant bacteria under the high temperature conditions of Daqu preparation. Bacillus can not only metabolize and produce flavor substances such as ACT, 2,3-butanediol and pyrazine...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12P7/26C12R1/07
CPCC12N1/20C12N1/02C12P7/26C12R2001/07C12N1/205
Inventor 罗惠波张玲玲赵东乔宗伟郑佳童文华黄丹杨莹
Owner WULIANGYE
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