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Isolated culture method of caproic acid bacteria in pit mud

A technology for the isolation and cultivation of caproic acid bacteria, which is applied in the field of isolation and cultivation of microorganisms for wine making, can solve the problems of long time consumption and difficulty in screening high-quality caproic acid bacteria, so as to accelerate aging, strengthen the ability of caproic acid production, and increase the rate of high-quality wine Effect

Inactive Publication Date: 2020-12-18
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Most of the microorganisms in the pit mud are non-cultivable microorganisms, which makes the screening of high-quality caproic acid bacteria very difficult, and the traditional method of obtaining caproic acid bacteria takes too long

Method used

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  • Isolated culture method of caproic acid bacteria in pit mud

Examples

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Comparison scheme
Effect test

Embodiment 1

[0037] Screening of caproic acid bacteria from 5-year-old pit mud of a winery in Sichuan:

[0038] (1) Preparation of pit mud suspension

[0039] Weigh 2.5g of pit mud, add it into a conical flask filled with 50mL sterile normal saline and 20 sterile glass beads (φ=3mm), shake at 37°C and 220r / min for 30min;

[0040] (2) Enrichment culture of caproic acid bacteria in pit mud

[0041] Inoculate the pit mud suspension with a volume fraction of 3% in 300 mL of RCM liquid medium, and enrich and culture in an anaerobic incubator at 37°C for 4 days.

[0042] (3) Electric field assisted primary screening

[0043] Pour 200mL of the enriched cultured liquid medium into the electric field-assisted screening device, apply a voltage of 8V between the two electrode plates, change the applied voltage to 12V after culturing for 0.5h, and change the applied voltage to 14v after cultivating for 1h. Obvious material attachment is observed on the board and the voltage is stopped;

[0044] (4...

Embodiment 2

[0049] Screening of caproic acid bacteria from 15-year-old pit mud of a winery in Sichuan:

[0050] (1) Preparation of pit mud suspension

[0051] Weigh 2.5g of pit mud, add it into a conical flask filled with 50mL sterile normal saline and 20 sterile glass beads (φ=3mm), shake at 37°C and 220r / min for 30min;

[0052] (2) Enrichment culture of caproic acid bacteria in pit mud

[0053] Inoculate the pit mud suspension with a volume fraction of 3% in 300 mL of RCM liquid medium, and enrich and culture in an anaerobic incubator at 37°C for 4 days.

[0054] (3) Electric field assisted primary screening

[0055] Pour 200mL of the enriched liquid culture medium into the electric field-assisted screening device, apply a voltage of 6V between the two electrode plates, and change the applied voltage to 10V after 0.5h of cultivation, and change the applied voltage to 13v after culturing for 0.5h. Obvious substance attachment was observed on the negative plate and the voltage was stop...

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Abstract

The invention provides an isolated culture method of caproic acid bacteria in pit mud. The isolated culture method comprises the following steps of carrying out enrichment culture by using an RCM liquid culture medium, and performing isolated culture on caproic acid bacteria in Luzhou-flavor liquor pit mud by using an external electric field, thereby shortening the acquisition time of caproic acidbacteria, and enhancing the caproic acid production capacity of caproic acid bacteria. The method can meet the requirement for isolated culture of microorganisms in the research direction of white spirit brewing, vinegar brewing, fermented food and the like, the yield of caproic acid in the caproic acid bacteria fermentation liquor obtained through the method can reach 15-18 mg / mL, the method hasthe advantages of being simple, rapid and efficient, isolated culture time of caproic acid bacteria is greatly shortened, production efficiency is improved, and aging of pit mud is accelerated, and the high-quality wine rate of a manufacturer is improved.

Description

technical field [0001] The invention relates to a method for isolating and cultivating microorganisms for brewing wine, in particular to a method for isolating and cultivating caproic acid bacteria in pit mud. Background technique [0002] my country has a long history of wine culture. Liquor occupies a dominant position in alcoholic beverages. Luzhou-flavor liquor accounts for more than 70% of the liquor market. The quality of Luzhou-flavor liquor is closely related to the quality of pit mud. Both the construction of new cellars and the maintenance of old cellars require large quantities of pit mud. The quality of pit mud seriously affects the quality of Luzhou-flavor liquor. After many years of research on pit mud microorganisms, artificial pit mud technology has achieved certain development. At present, the well-applied pit mud production technology is to add various enhanced functional bacteria to the pit mud in the process of making pit mud, so as to achieve the purpo...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12N13/00
CPCC12N1/02C12N13/00
Inventor 张朝正赵华刘安张天爽
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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