Mutant strain for high yield of gamma-aminobutyric acid and biological agent of mutant strain

A technology for mutagenizing strains and aminobutyric acid, applied in the directions of microorganism-based methods, microorganisms, microorganisms, etc., can solve the problems of increasing product production costs, detrimental to body health, and affecting food flavor and taste.

Active Publication Date: 2019-01-11
湖南肯基因科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 r

Method used

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  • Mutant strain for high yield of gamma-aminobutyric acid and biological agent of mutant strain
  • Mutant strain for high yield of gamma-aminobutyric acid and biological agent of mutant strain
  • Mutant strain for high yield of gamma-aminobutyric acid and biological agent of mutant strain

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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 mutant lactobacillus plantarum and a biological agent of the lactobacillus plantarum. The mutant strain is KJY-HN001-01-02 having a collection number of CGMCC NO. 15422, and thecollection date is March 07, 2018. The invention further provides a biological agent prepared from the mutant strain, and an application used in high yield of gamma-aminobutyric acid. The invention further provides an application of the mutant strain alone or combined with lactic acid bacteria for complexing so as to produce sour soybean milk, yoghurt tofu, bean liquor drink and bean liquor soursoup enriched in gamma-aminobutyric acid.

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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IPC IPC(8): C12N1/20C12N15/01C12P13/00A23C20/02A23C11/10C12R1/25A23L11/45A23L11/50A23L11/60
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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