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Ethanol-tolerant lactobacillus plantarum and application thereof in fermented food

A technology of Lactobacillus plantarum and ethanol resistance, applied in the field of microorganisms, can solve the problems that lactic acid bacteria are not suitable for high ethanol alcohol fermentation process, etc., and achieve the effect of improving sensory quality, controllable production process and simple operation

Active Publication Date: 2020-04-14
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-concentration ethanol is the source of the ability to form rice wine, fruit wine, rice wine and apple cider vinegar. However, common lactic acid bacteria are not suitable for high-ethanol alcohol fermentation. Therefore, a strain of Lactobacillus plantarum that is resistant to ethanol and high in GABA production is provided appears to be particularly important

Method used

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  • Ethanol-tolerant lactobacillus plantarum and application thereof in fermented food
  • Ethanol-tolerant lactobacillus plantarum and application thereof in fermented food
  • Ethanol-tolerant lactobacillus plantarum and application thereof in fermented food

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1: Obtaining of ethanol-resistant and high-yielding γ-aminobutyric acid Lactobacillus plantarum XH-1 and Lactobacillus plantarum F of the present invention

[0025] Weigh 10g of wine mash and add them into conical flasks filled with 90mL of sterile water, add sterilized glass beads, shake fully on a vortex shaker for 10min, so that the wine mash is fully dispersed. The supernatant was subjected to a 10-fold serial dilution, and the dilution was 10 -3 、10 -4 、10 -5 、10 -6 、10 -7 , use a pipette gun to draw 0.5mL sample solution, and use the plate coating method to evenly spread the sample solution on the MRS screening medium with different ethanol concentrations, and place the prepared plate upside down in a constant temperature incubator at 36°C for 5 days. Observe and record the characteristic morphology of bacterial colonies. Pick the bacterial strains that can grow well on the medium with 12% and 17% ethanol additions and isolate a single colony through ...

Embodiment 2

[0026] Example 2: Comparison of ethanol-resistant and high-yielding γ-aminobutyric acid Lactobacillus plantarum XH-1 and Lactobacillus plantarum F in the MRS system with different concentrations of ethanol

[0027] From Figure 4 It can be seen that the OD600nm of Lactobacillus plantarum XH-1 reached 4.42 in the MRS liquid medium added with 15% ethanol, and the OD600nm reached 3.82 under 17% ethanol, showing good growth. However, the OD600nm of Lactobacillus plantarum F was only 0.21 and 0.18 under the MRS liquid medium added with 15% and 17% ethanol, and the growth was severely inhibited by high concentration of ethanol, and basically could not grow out.

Embodiment 3

[0028] Example 3: 16S rRNA identification experiment of Lactobacillus plantarum XH-1 with ethanol resistance and high γ-aminobutyric acid production in the present invention

[0029] The 16S rDNA gene method was used to identify the isolated and screened bacteria in molecular biology. According to the highly conserved 16S rDNA gene sequence of prokaryotic organisms, universal primers are designed, and the 16S rDNA gene fragment of the bacteria is amplified using the DNA of the isolate as a template, and the 16S rDNA gene sequence of the isolate is determined, and homologous to the gene sequence in GenBank Sexual comparison, in order to determine the species of isolates.

[0030] The whole genome DNA of ethanol-resistant Lactobacillus plantarum XH-1 (Lactobacillus plantarum XH-1) was extracted with the bacterial genomic DNA rapid extraction kit, and PCR amplification was carried out with bacterial 16S rDNA universal primers. PCR amplification system (25 μL): 10×PCR Buffer 2.5μ...

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Abstract

The invention discloses an ethanol-tolerant lactobacillus plantarum and application thereof in fermented food. The lactobacillus plantarum XH-1 is classified and named as Lactobacillus plantarum and is preserved in China General Microbiological Culture Collection Center on August 16, 2019. The preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the preservationnumber is CGMCC NO.18390. The alcohol tolerance of the ethanol-tolerant lactobacillus plantarum disclosed by the invention can reach 17% (V / V), the ethanol-tolerant lactobacillus plantarum has the characteristic of high yield of gamma-aminobutyric acid, the capacity of producing gamma-aminobutyric acid in an MRS culture medium reaches 170g / l, and the ethanol-tolerant lactobacillus plantarum is applied to wine and vinegar brewing production. The strain can normally grow and reproduce and produce gamma-aminobutyric acid at high yield in a high-concentration ethanol environment for wine and vinegar brewing, the production process is controllable, and the operation is simple and convenient; the content of lactic acid, ethyl lactate and phenethyl alcohol in the finished wine and vinegar is increased, and the sensory quality of the finished wine and vinegar can be greatly improved.

Description

technical field [0001] The invention relates to an ethanol-resistant plant lactobacillus and its application in fermented food, belonging to the technical field of microbes. Background technique [0002] Rice wine is a low-alcohol drinking wine, its alcohol content is generally 3% to 14%, and it belongs to low-alcohol fermented wine. Modern nutrition thinks that the nutritional value of rice wine is that except water and ethanol, the wine liquid also contains nutrients such as sugar, organic acids, proteins, amino acids, vitamins and minerals. Due to the existence of these functional substances, rice wine has effects such as promoting blood circulation, promoting metabolism, and prolonging life. Fruit wine is a low-alcohol drink made from fruit through fermentation. It is rich in sugar, organic acids, esters and multivitamins, and rich in SOD, which can enhance immune function. Alcohol products have a very broad market. Fruit wine is the development direction of the wine i...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12J1/04C12G3/02C12R1/25
CPCC12G3/02C12J1/04C12N1/20C12N1/205C12R2001/25
Inventor 许正宏张晓娟解寒史劲松陆震鸣柴丽娟彭铭烨邓永建刘瑞山
Owner JIANGNAN UNIV
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