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A mutant strain with high γ-aminobutyric acid and its biological preparation

A technology of aminobutyric acid and mutagenic strains, applied in microorganism-based methods, microorganisms, microorganisms, etc., can solve the problems of restricting application prospects, high glutamic acid intake, and high production costs

Active Publication Date: 2021-06-29
湖南肯基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this production method requires the selection of special ganoderma lucidum fermentation broth components, the addition of high-concentration substrates, and the need to repeatedly explore new fermentation optimization factors, resulting in high production costs and excessive complexity of the process, which restricts industrialization. Application prospects
[0016] In addition, most of the reported lactic acid bacteria capable of producing GABA need to use high-concentration glutamic acid or its salts as substrates to ensure the production of GABA. This high-concentration substrate not only affects the flavor and taste of related foods, It also increases the production cost of the product, and at the same time, a large intake of glutamic acid is not conducive to the health of the body

Method used

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  • A mutant strain with high γ-aminobutyric acid and its biological preparation
  • A mutant strain with high γ-aminobutyric acid and its biological preparation
  • A mutant strain with high γ-aminobutyric acid and its biological preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Embodiment 1, experimental material

[0086] Starting strain:

[0087] 1. Lactobacillus plantarum is the frozen strain Lp20160820 of our company.

[0088] 2. Lactobacillus rhamnosus: selected from the mutagenic strain Lactobacillus rhamnosus KJY11, the preservation number is CGMCC No.15421, and the preservation date is March 07, 2018.

[0089] 3. Lactobacillus zea: selected from the mutagenic strain Lactobacillus zea KJY13, the preservation number is CGMCC No.15423, and the preservation date is March 07, 2018.

[0090] 4. Lactobacillus casei: selected from the mutagenized strain Lactobacillus casei (Lactobacillus casei strain) KJY14, the preservation number is CGMCC No.15424, and the preservation date is March 07, 2018.

[0091] 5. Leuconostoc mesenteroides strain: selected from the mutagenic strain Leuconostoc mesenteroidesstrain KJY15, the preservation number is CGMCC No.15425, and the preservation date is March 07, 2018.

[0092] 6. MRS liquid medium: peptone 10g / ...

Embodiment 2

[0097] Embodiment 2, microwave mutagenesis Lactobacillus plantarum

[0098] The frozen Lactobacillus plantarum strains were respectively activated twice in the MRS liquid medium, and then spread on the MRS solid medium by the concentration gradient method. Anaerobic culture was carried out at 37°C for 20-30 hours, and a single colony with obvious calcium dissolution circle was selected for streak purification after plate counting. The final single colonies obtained were purified for 3 generations.

[0099] Collect strains and dilute to 10 7 / ml of bacterial suspension, placed in a microwave oven with a power of 500W, and the irradiation time was 10, 20, 30, 40, 50, 60, 70, 80, 90s, respectively. Take it out every 10s and use an ice bath for 10s to eliminate the thermal effect of the microwave. After refrigerated at 4°C for 12h in the dark, spread on the MRS solid medium plate, incubate at 37°C for 24h, count the colonies, and calculate the lethality.

[0100] Using 500 powe...

Embodiment 3

[0107] Embodiment 3, nitrosoguanidine mutagenesis

[0108] Nitrosoguanidine (NTG): Take 0.1g of nitrosoguanidine and add 10mL of acetone as co-solvent, after complete dissolution, take 1mL of nitrosoguanidine acetone solution and add 9mL of sodium phosphate buffer (pH7.4, 0.02mol / L) to prepare Prepare a nitrosoguanidine stock solution with an NTG concentration of 1 mg / mL.

[0109] The cultivated Lactobacillus plantarum was collected and diluted to 10 8 / ml of bacterial suspension. Then take 8 mL of bacterial suspension, add 2 mL of nitrosoguanidine stock solution, and the final concentration of NTG is 200 μg / mL, and set the non-mutated stock solution as a control.

[0110] Put the above-mentioned bacterial solution in a conical flask, incubate on a shaking table at 37°C in the dark for 10, 15, 20, 25, 30, 35, 40, 45, and 50 min, then add physiological saline to terminate the reaction; After the bacterial solution was ice-bathed for 2-3 hours to induce the positive mutation,...

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Abstract

The invention provides a mutagenized Lactobacillus plantarum and its biological preparation, wherein the mutagenized bacteria is (KJY‑HN001‑01‑02), CGMCCNO.15422, and the preservation date is March 07, 2018. The present invention also provides a biological preparation prepared by the mutagenic bacteria for high-yielding γ-aminobutyric acid. The present invention also provides the use of the mutagenic bacteria alone or in combination with other lactic acid bacteria to produce yogurt rich in γ-aminobutyric acid, yogurt tofu, bean clear drink, and bean clear sour soup.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to a mutagenized Lactobacillus plantarum, and uses the mutagenized bacteria to high-yield gamma-aminobutyric acid. Background technique [0002] Gamma‐aminobutyric acid (GABA), also known as aminobutyric acid and 4‐aminobutyric acid, has a molecular formula of C4H9O2N and a relative molecular mass of 103.12. The structural formula and three-dimensional structure are as follows: [0003] [0004] γ-aminobutyric acid is a new resource food approved by the National Health and Family Planning Commission in 2017. γ-aminobutyric acid is a naturally occurring non-protein amino acid widely present in animals and plants. In plants, γ‐aminobutyric acid is involved in stress response and regulation of plant growth direction; in animals, γ‐aminobutyric acid is an important inhibitory neurotransmitter in the central nervous system and has extremely important physiological f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/01C12P13/00A23L11/45A23L11/65A23L11/50A23C9/123C12R1/25A23L11/60
CPCA23C11/106A23C20/025C12N1/20C12N15/01C12P13/005C12R2001/25C12N1/205A23V2400/169
Inventor 张胤李肯赵聃
Owner 湖南肯基因科技有限公司
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