Microbial complex microbial agent for degrading lactic acid

A technology of microbial preparations and microbial strains, applied in the field of microorganisms

Active Publication Date: 2020-06-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the domestic research on Pichia Kudriazweibi in liquor is still limited to the isolation and identification of a single strain and the study of the ferm

Method used

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  • Microbial complex microbial agent for degrading lactic acid
  • Microbial complex microbial agent for degrading lactic acid
  • Microbial complex microbial agent for degrading lactic acid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The screening of embodiment 1 bacterial strain

[0038] During the fermentation process of Maotai-flavored liquor, a strain of lactic acid-tolerant Pichia kudriazwii was screened, and the process was as follows.

[0039] Samples of fermented grains of a Maotai-flavor liquor were collected. Weigh 10g of sample into a 250mL Erlenmeyer flask, add 90mL sterile saline, add glass beads and shake evenly. Aspirate 0.1mL supernatant into 100mL enriched medium, culture at 30°C, 200rpm for 2-4 days, observe whether the culture medium is turbid; if the culture medium is obviously turbid, absorb 0.1mL enriched culture medium In 100mL liquid screening medium, culture at 30°C and 200rpm for 2 days. After 3 to 4 times of culture, the culture solution was gradually diluted on YPD solid medium. After 3 to 4 days of culture, the single colony was the target strain with anti-lactic acid properties.

[0040] Randomly pick 7 single bacterium colonies from this plate, inoculate until adding...

Embodiment 2

[0041] The identification of embodiment 2 bacterial strains

[0042] (1) Colony characteristics and cell morphology

[0043] The colony is white, with rough surface and uniform texture, easy to pick. As a result of observing the morphology through a microscope, it was found that the isolated and screened strain cells were elliptical, and some of them were budding and dividing.

[0044] (2) Physiological and biochemical characteristics

[0045] DC-16 was inoculated in sorghum juice medium, and after 1 day of fermentation at 30°C and 200rpm, DC-16 entered a stationary phase. Such as Figure 4 As shown, when fermented for 3 days, the biomass OD of DC-16 600 Up to 4.48.

[0046] The glucose content in the sorghum juice medium is 58g·L -1 . Such as Figure 5 As shown, after 60 hours of fermentation, DC-16 can convert 58g·L of sorghum juice medium -1 The glucose is completely consumed and produces 13.06g·L -1 ethanol.

[0047] (3) Molecular biological identification

[0...

Embodiment 3

[0050] Embodiment 3 Yeast of the present invention is to the degradation situation of lactic acid

[0051] Using sorghum juice medium with different lactic acid concentrations as the fermentation medium, the experimental strain Pichia kudri Azwiya DC-16 (hereinafter referred to as P.k.DC-16) and its model strain Pichia kudri Azwiya Yeast ATCC24210 (hereinafter referred to as P.k.ATCC 24210) was used for lactic acid metabolism experiments. According to the detection results of lactic acid in fermented grains during the screening process, the addition amount of lactic acid in the metabolic experiment was designed to be 20, 30, and 40 g·L respectively. -1 .

[0052] Experimental and control group settings:

[0053] Control group 1: add 20g·L to -1 Lactic acid sorghum juice medium was inoculated with P.k.ATCC24210 at 7% inoculation volume (v:v);

[0054] Experimental group 1: add 20g·L to -1 Inoculate P.k.DC-16 in lactic acid sorghum juice medium at 7% inoculation volume (v:v...

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Abstract

The invention discloses a microbial complex microbial agent for degrading lactic acid, and belongs to the technical field of microbes. The degradability of the pichia kudriavzevii provided by the invention to the lactic acid is up to 12.69 g/L, and is 2.04 times that of a model strain; at the same time, the strain can metabolize ethanol, and the OD600 can reach 4.48 after the strain is fermented in a sorghum juice culture medium at 30 DEG C and 200 rpm for 3 days; after fermentation is performed for 60 h, 58 g/L glucose in the sorghum juice culture medium can be completely depleted, and 13.06g/L ethanol can be produced; and the pichia kudriavzevii can relieve lactic acid pressure in a fermentation system when degrading the lactic acid, so that saccharomyces cerevisiae can grow and be metabolized to produce liquor.

Description

technical field [0001] The invention relates to a microbial composite bacterial agent for degrading lactic acid, which belongs to the technical field of microorganisms. Background technique [0002] Liquor is a typical example of traditional Chinese fermented food. During the anaerobic fermentation process, acid-producing microorganisms such as lactic acid bacteria metabolize to produce a large amount of acid substances, which makes the non-volatile acid substances in the wine grains continuously accumulate. Lactic acid is the main organic acid in the fermented grains, and the content in the fermented grains of Maotai-flavored liquor reaches 20-40g·kg -1 Fermented grains, the content of fermented grains in Luzhou-flavor liquor reaches 20-30g·kg -1 Wine grains. The accumulation of lactic acid leads to an increase in the acidity of the fermented grains of liquor, causing microbial cells to stop growing and even die. The undissociated form of lactate is lipophilic, so lactat...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N1/18C12G3/02C12R1/84C12R1/865
CPCC12N1/16C12N1/18C12G3/02C12R2001/84C12N1/165C12G1/0203C12R2001/865C12N1/185C12P39/00A01P3/00A01N63/32A23B9/26A23L3/3571C12G3/021A21D8/047A23C19/0325A23L27/50C12G3/022A01P1/00A23V2002/00C12G2200/05
Inventor 杜海徐岩邓楠
Owner JIANGNAN UNIV
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