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Clostridium tyrobutyricum for reducing content of n-butyl alcohol in Baijiu fermentation process

A technology of clostridium tyrobutyricum and starter, which is applied in the field of clostridium tyrobutyricum, can solve the problems of uncontrollable influence on the safety of wine body and flavor, and achieve the effect of reducing and controlling the content of n-butanol and increasing the amount of production

Active Publication Date: 2021-04-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Genetic modification of microorganisms to regulate n-butanol metabolism can block its production pathway from the source, which is currently the most effective method. produce uncontrollable effects; therefore, screening strains that can reduce the production of n-butanol during liquor fermentation is currently the safest and most effective method

Method used

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  • Clostridium tyrobutyricum for reducing content of n-butyl alcohol in Baijiu fermentation process
  • Clostridium tyrobutyricum for reducing content of n-butyl alcohol in Baijiu fermentation process
  • Clostridium tyrobutyricum for reducing content of n-butyl alcohol in Baijiu fermentation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Isolation and Identification of Pit Mud Microorganisms

[0034] Weigh 5g of pit mud sample into 100mL RCM liquid medium with glass beads, shake well, heat-treat at 80°C for 10min to eliminate non-spore bacteria, and anaerobically culture at 37°C for 4-7d; The supernatant was centrifuged at 4°C and 12,000rpm for 5min, and the supernatant was taken; the supernatant was serially diluted for 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 , absorb 0.1mL of each gradient bacterial solution and spread it on the RCM medium, and culture it anaerobically at 37°C for 2-5 days; pick colonies of different shapes to streak on the RCM solid medium, and then culture them anaerobically at 37°C 4-7 days until colonies grow; follow the above steps to streak and isolate 3 times to obtain single colonies of each strain.

[0035] Pick a single colony on the plate and inoculate it into 8-10mL RCM liquid medium, and culture it anaerobically at 37°C for 24-28 hours to obtain the bacterial so...

Embodiment 2

[0040] Example 2: Screening for reducing n-butanol production strains of Clostridium beijerinckii 6Y-1

[0041] Introduce the strains of the identified strains into RCM liquid medium for anaerobic culture at 37°C for 24-28h to OD 600 1.1, each strain was mixed with Clostridium beijerinckii 6Y-1 with the strongest n-butanol synthesis ability at an inoculation ratio of 10:1 by volume, and then inserted into the fermentation medium. culture medium. All were inoculated into the butanol fermentation medium with an inoculum amount (after mixing) of 10% (10 mL / 100 mL) of the volume of the butanol medium, and cultured anaerobically at 37° C. for 3 days. An internal standard (tert-amyl alcohol with a final concentration of 10 mg / L) was added to the fermentation broth, mixed evenly and then tested.

[0042] Compared with only adding Clostridium bejejerinckii 6Y-1 control, adding Clostridium tyrobutyricum ZY-4 can reduce the content of n-butanol by 68.91% ( figure 1 ).

Embodiment 3

[0043] Example 3: Analysis of the reduction and control mechanism of Clostridium tyrobutyricum ZY-4 in the butanol medium system

[0044] Clostridium tyrobutyricum ZY-4 with a reduction effect and high butanol-producing Clostridium beijerinckii 6Y-1 were mixed at an inoculation ratio of 10:1 by volume and then inserted into the butanol fermentation medium, and the butanol medium After the inoculum amount of 10% of the volume was inoculated, cultured anaerobically at 37°C for 3 days. The butanol content and by-products, including acetic acid, butyric acid, ethanol and acetone, generated from glucose, an important precursor, were determined by gas chromatography and high performance liquid chromatography, respectively.

[0045] The study of the downregulation mechanism was carried out in the butanol medium with glucose as the precursor. Compared with the control that only added Clostridium beijerinckii 6Y-1, the addition of Clostridium tyrobutyricum ZY-4 could reduce the produc...

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Abstract

The invention discloses clostridium tyrobutyricum for reducing the content of n-butyl alcohol in a Baijiu fermentation process. According to the invention, strains are screened and separated from pitmud, the clostridium tyrobutyricum for reducing the content of the n-butyl alcohol is obtained through screening in a butanol culture medium, then influence of different addition proportions on synthesis of the n-butyl alcohol during co-culture is investigated in a five-grain culture medium system, and finally, a pit mud fermentation system is simulated to verify the reduction and control effect of the clostridium tyrobutyricum. In the simulated fermentation process of Baijiu, compared with a blank control only containing the pit mud, the clostridium tyrobutyricum is added, so that the generation amount of the n-butyl alcohol in the fermentation process can be reduced by 30.05%, and the content of a skeleton flavor substance namely ethyl caproate of strong aromatic Baijiu is increased by 25.77%; Compared with a high-butanol-yield fermentation system with clostridium beijerinckii 6Y-1, the system with the clostridium tyrobutyricum has the advantages that the generation amount of the n-butyl alcohol in the system can be reduced by 27.08%, and the content of the ethyl caproate is increased by 5.09%.

Description

technical field [0001] The invention relates to a Clostridium tyrobutyricum strain for reducing n-butanol content in the fermentation process of liquor, belonging to the technical field of bioengineering. Background technique [0002] Higher alcohol (resin oil alcohol) refers to monohydric alcohols with 3 carbon atoms or more, which is a restrictive index in the hygienic standards of alcoholic products. Appropriate content of high-grade alcohol can improve the richness of the wine and increase the coordination of the wine. If there is little or no higher alcohol, the wine will have a weak taste; but too much content will cause headaches, dizziness, etc., and it is also one of the main reasons for getting drunk. N-butanol is one of the main components of higher alcohols, therefore, its content in liquor needs to be reduced to an appropriate concentration. [0003] Mainly control n-butanol in liquor from two aspects at present: 1, improve fermentation process. The content o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12G3/022C12G3/021C12R1/145
CPCC12G3/022C12G3/021Y02E50/10
Inventor 方芳勾文君王芊婷吴浪涛谢专丁文骏陈坚堵国成
Owner JIANGNAN UNIV
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