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Lactobacillus casei stain mutant strain for producing glutamine transaminase and use thereof

A kind of Lactobacillus casei, mutagenic strain technology, applied in the field of biological preparations, can solve the problem of failing to accurately measure the size of glutamine aminotransferase enzyme activity, not involving and the like

Active Publication Date: 2019-01-25
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

[0005] In 2001, Lu Zhaoxin of Nanjing Agricultural University and his collaborators screened Streptomyces griseignis producing TGase from the soil, but failed to accurately determine the enzyme activity of the strain producing glutamine transaminase
These methods either add TG enzyme / or chymosin alone, or play the probiotic effect of Lactobacillus casei, and do not directly use Lactobacillus casei to produce TG enzyme

Method used

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  • Lactobacillus casei stain mutant strain for producing glutamine transaminase and use thereof
  • Lactobacillus casei stain mutant strain for producing glutamine transaminase and use thereof
  • Lactobacillus casei stain mutant strain for producing glutamine transaminase and use thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, experimental material

[0065] Starting strain:

[0066] 1. Lactobacillus casei is the frozen strain LC20170301 of our company.

[0067] 2. Lactobacillus rhamnosus mutagenized strain KJY11, the preservation number is CGMCC No.15421, and the preservation date is March 07, 2018.

[0068] 3. The Lactobacillus plantarum mutagenic strain KJY12, the preservation number is CGMCC No.15422, and the preservation date is March 07, 2018.

[0069] 4. The Lactobacillus zea mutagen strain KJY13, the preservation number is CGMCC No.15423, and the preservation date is March 07, 2018.

[0070] 5. Leuconostoc enterococcus mutagen strain KJY15, the preservation number is CGMCC No.15425, and the preservation date is March 07, 2018.

Embodiment 2

[0071] Embodiment 2, microwave mutagenesis Lactobacillus casei

[0072] Collect the domesticated 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.

[0073] Using 500 power to irradiate the bacterial suspension of Lactobacillus, the relationship between the lethality and the irradiation time is as follows figure 1 shown.

[0074] Lethal rate (%) = (total bacterial count without mutagenization treatment - surviving bacterial count after mutagenization treatment) / total bacterial count without mutagenization treatment × 100

[0075] It can be seen that with the increase of microwave time, t...

Embodiment 5

[0080] Embodiment 5, nitrosoguanidine mutagenesis

[0081] 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.

[0082] Collect the cultured acclimated strains and dilute 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.

[0083] Put the above-mentioned bacterial solution in a Erlenmeyer flask, shake on a shaking table at 37°C and incubate in the dark for 10, 15, 20, 25, 30, 35, 40, 45, and 50 min, respectively, and then add physiological saline to terminate the reaction; After the bacterial solution was ice-bathed for 2-3 hours to induce the p...

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Abstract

The invention provides a mutagenic Lactobacillus casei strain and a biological preparation thereof. The mutant strain is Lactobacillus casei strain KJY14(KJY-HN001-01-04), CGMCC No.15424, deposited onMarch 7, 2018. The invention also provides a biological preparation containing Glutamine transaminase prepared by the mutagenic bacterium. The invention also provides the use of the mutant strain alone or in combination with other lactic acid bacteria for preparing coagulant for soy yoghurt, yoghurt tofu, soybean albumen beverage and soybean albumen acid soup.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to a mutagenized Lactobacillus casei, which is used to produce transglutaminase, and which is used to prepare biological preparations for coagulation through the mutagenic bacteria. use. Background technique [0002] Transglutaminase (Transglutaminase, E.C.2.3.2.13), referred to as TGase, is a biological enzyme with functional properties such as improving protein structure, flavor and shelf life. TG enzyme is mainly derived from the liver, blood, hair follicle and fur of mammals; it is mainly produced by microorganisms in industrial production; in addition, the enzyme has also been found in some plant tissues. According to different sources, TG enzymes are generally divided into tissue transglutaminase (Tissue Transglutaminase, tTG) and microbial transglutaminase (Microbial Transglutaminase, MTG). Transglutaminase was first isolated from the liver of guinea pigs ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N9/10A23L29/269A23L2/52A23C9/154A23C11/10C12R1/245A23L11/45A23L11/50A23L11/65
CPCA23C9/1544A23C11/103A23L2/52A23L29/269C12N9/1044C12N15/01C12Y203/02013C12R2001/245C12N1/205A23V2400/161A23V2400/125A23V2400/187A23V2400/175A23V2400/169
Inventor 张胤李肯赵聃
Owner 湖南肯基因科技有限公司
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