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Lactobacillus rhamnosus and application thereof

A technology for preservation of Lactobacillus rhamnosus and strains, which is applied in the field of probiotics and achieves the effects of excellent simulated gastrointestinal fluid tolerance and inhibition of Escherichia coli

Active Publication Date: 2021-11-05
HUNAN AGRICULTURAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the means of treating such diseases are drug treatments, but drug treatments have certain side effects, so effective and safe treatments need to be discovered

Method used

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  • Lactobacillus rhamnosus and application thereof
  • Lactobacillus rhamnosus and application thereof
  • Lactobacillus rhamnosus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Isolation and purification of strains

[0041]1) Sample collection: Sampling and numbering were carried out in 50mL sterile centrifuge tubes in Nanshan pasture (different milk stations), Wangcheng (dairy company and pasture) and Ningxiang (dairy company and pasture) in Hunan Province respectively, and used a sampling box containing ice packs Rapid transport to microbiology laboratory; a total of 71 samples were collected from these areas.

[0042] 2) Isolation of lactic acid bacteria: under aseptic conditions, 25 g of the sample was taken and mixed in a 500 mL Erlenmeyer flask containing 225 mL of sterile normal saline (0.85% NaCl), and diluted to 10 with a 10-fold gradient. -2 ~10 -6 , and in the MRS solid (containing 0.5% CaCO 3 (w / v)) were coated on the plate, and cultured at 37° C. for 48 h. Pick a single colony with a transparent calcilytic circle for Gram staining, and purify the positive strains by streaking three times and save them by numbering. All the...

Embodiment 2

[0082] Example 2 In vivo evaluation of bacterial count antiallergic activity in mice

[0083] (1) Activation of the strain

[0084] After activating the original strain of Lactobacillus rhamnosus AU9260 stored in a -80°C refrigerator for 2 generations, streak it on the solid MRS medium, culture it statically at 37°C for 72 hours, and then pick a single colony and inoculate it in the MRS liquid medium , after constant temperature cultivation at 37°C for 24 hours, inoculate into liquid MRS medium according to 1% (v / v) inoculum amount, and continuously culture for 2 generations.

[0085] (2) Preparation of bacterial suspension

[0086] The activated Lactobacillus rhamnosus AU9260 was inoculated in MRS liquid medium, cultivated at 37°C for 24 hours, and then centrifuged at 8 000 r / min for 4 minutes to collect the bacterial pellet, then resuspended in PBS buffer, and adjusted the bacterial concentration to 10 8 CFU / mL, 10 10 CFU / mL.

[0087] (3) Construction of animal models

...

Embodiment 3

[0130] Example 3 Effect of β-lactoglobulin-sensitized mice on intestinal flora

[0131] (1) Establishment of mouse model

[0132] The mouse models in this example were established according to Example 2.

[0133] (2) DNA extraction from mouse feces samples

[0134] The fresh feces of mice in each group were collected, frozen in time at -80°C, and genomic DNA was extracted using a DNA extraction kit according to the instructions.

[0135] (3) PCR amplification

[0136] The DNA extracted in the previous step was used as a template for amplification, and the amplified region was the V3-V4 region of the bacterial 16S rRNA gene. Primer sequences, amplification systems, and amplification conditions are shown in Tables 7, 8, and 9.

[0137] Table 7 Primer Sequence

[0138]

[0139] Table 8 Amplification system

[0140]

[0141]

[0142] Table 9 Amplification Conditions

[0143]

[0144] (4) Detection, merging and purification of PCR products

[0145] After the amp...

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Abstract

The invention discloses lactobacillus rhamnosus and an application thereof. The lactobacillus rhamnosus is named as lactobacillus rhamnosus AU9260 in taxonomy, and the preservation number of the lactobacillus rhamnosus is CGMCC No.21662. The strain has excellent in-vitro probiotic characteristics such as simulated gastrointestinal fluid tolerance, bacteriostatic activity, adhesion to Caco-2 cells and the like, and is preliminarily judged to be safe. In addition, the strain has a potential function of relieving and / or preventing allergic diseases of hosts, can significantly reduce histamine, specific IgE and cell factors in serum of mice induced and sensitized by beta-lactoglobulin, and can also protect damage to intestinal mucosa structures of the mice caused by allergy and regulate intestinal flora structures. The lactobacillus rhamnosus can be used as foods and / or health-care foods with a desensitization effect.

Description

technical field [0001] The invention belongs to the technical field of probiotics, and in particular relates to a kind of Lactobacillus rhamnosus and application thereof. Background technique [0002] Probiotics are defined as the general term for live beneficial microorganisms, supplementing probiotics in an appropriate amount in daily life is beneficial to the health of the host. Probiotics are microorganisms that can coexist harmoniously with humans. They can protect the health of the human host and ensure the orderly operation of the entire body in the body of the human host. Inhibit intestinal inflammation, improve host immunity, regulate intestinal microbiota and maintain structural balance, improve antioxidant levels and remove cholesterol, etc. Probiotics mainly include the following categories: Lactobacillus, Bifidobacterium, Streptococcus, yeast, etc. These bacteria not only can effectively protect and regulate the health of humans and animals, but also are widel...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/04A23C9/13A61K35/747A61P37/08C12R1/225
CPCC12N1/20C12N1/04A23C9/13A61K35/747A61P37/08A23V2400/175
Inventor 周辉汪家琦刘成国潘丽娜雷文平戴智勇康文丽吴忠坤李辉宇荣智兴
Owner HUNAN AGRICULTURAL UNIV
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