A strain of Lactobacillus fermentum capable of regulating intestinal tight junction proteins and its application

A Lactobacillus fermentum, compact technology, applied in the field of microorganisms, to achieve a wide range of application prospects, improve barrier function, and repair colonic mucosal damage

Active Publication Date: 2022-03-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no report that Lactobacillus fermentum can regulate the tight junction protein Claudin-1

Method used

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  • A strain of Lactobacillus fermentum capable of regulating intestinal tight junction proteins and its application
  • A strain of Lactobacillus fermentum capable of regulating intestinal tight junction proteins and its application
  • A strain of Lactobacillus fermentum capable of regulating intestinal tight junction proteins and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Isolation and screening of Lactobacillus fermentum CCFM1058

[0028] 1. Sample collection

[0029] The intestinal contents of the farm population in Changji, Xinjiang Province were collected. The samples were placed in stool tubes filled with 30% glycerin and stored in an insulated box with ice packs. After being brought back to the laboratory, they were quickly placed in a -80°C refrigerator for separation filter.

[0030] 2. Isolation and purification of lactic acid bacteria

[0031] (1) Dilution coating: take about 0.5 g of the content preserved in 30% glycerol and add it into a 10 mL centrifuge tube containing 4.5 mL of normal saline under a sterile environment to obtain 10 -1Diluent, repeat the above dilution steps to obtain 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Diluent;

[0032] (2) Coating culture: pipette 100 μL of the above 10 -4 、10 -5 、10 -6 The three gradient dilutions were separated and screened in the human intestinal tract on the LFMATA ...

Embodiment 2

[0048] Example 2: Regulatory effect of Lactobacillus fermentum CCFM1058 on tight junction protein Claudin-1

[0049] 1. Preparation of gastric administration solution of Lactobacillus fermentum CCFM1058

[0050] The secondary purified culture solution of Lactobacillus fermentum CCFM1058 obtained in Example 1 was inoculated in the MRS solid medium with an inoculum size of 2% to 5% (v / v), and cultivated under anaerobic conditions at 37°C for 36-48h to obtain Single colony; pick a single colony on the MRS solid medium and inoculate it in the MRS medium, enrich and culture it under anaerobic conditions at 37°C for 16-18 hours for activation, and activate two generations continuously to obtain an activated culture; Inoculate 2% to 5% (v / v) of the inoculum in MRS liquid medium, enrich and culture under anaerobic conditions at 37°C for 16 to 18 hours to obtain bacterial liquid, centrifuge at 6000g for 10 minutes to obtain bacterial sludge, and use sterile saline After washing for 3 ...

Embodiment 3

[0079] Embodiment 3: the application of Lactobacillus fermentum CCFM1058

[0080] Lactobacillus fermentum CCFM1058 can be used to prepare milk, and the specific preparation process of milk is as follows:

[0081] (1) Inoculate the secondary purified culture solution of Lactobacillus fermentum CCFM1058 obtained in Example 1 into the culture medium with an inoculum size of 3% (v / v), and cultivate it at a temperature of 37° C. for 18 hours to obtain a bacterial solution; centrifuged to obtain the bacteria sludge; the bacteria sludge was washed 3 times with pH 7.2 phosphate buffer solution and then resuspended with a protective agent to a concentration of 1×10 10 CFU / mL to obtain a suspension; the suspension was pre-incubated at a temperature of 37°C for 60 minutes and then freeze-dried to obtain a starter;

[0082] Among them, the preparation method of the culture medium is: use 87.7% water based on the total weight of the culture medium to dissolve 10% enzyme-hydrolyzed skim mi...

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Abstract

The invention discloses a lactobacillus fermentum capable of regulating intestinal tight junction proteins and application thereof, belonging to the technical field of microbes. Lactobacillus fermentum CCFM1058 provided by the present invention can effectively improve the barrier function of intestinal mucosa and increase the expression of tight junction protein Claudin-1 (compared with the colitis group, Lactobacillus fermentum CCFM1058 has improved the expression level of Claudin-1 protein gene by nearly double), repair colonic mucosal damage. Lactobacillus fermentum CCFM1058 increased the level of tight junction protein Claudin-1 significantly better than the model strain Lactobacillus fermentum strain CECT5716. Therefore, the Lactobacillus fermentum strain that regulates the tight junction protein Claudin‑1 to protect the intestinal mucosal barrier of the present invention has broad application prospects in the direction of food and microecological preparations.

Description

technical field [0001] The invention relates to a lactobacillus fermentum capable of regulating intestinal tight junction proteins and application thereof, belonging to the technical field of microbes. Background technique [0002] The mucosal barrier is the most important barrier in the intestinal tract. It is composed of intact intestinal epithelial cells and the junctions between adjacent intestinal epithelial cells. It has selective permeability and regulates the transepithelial transport of water and solutes (such as simple sugar , amino acids, vitamins and hormones, etc.). Various connection structures between intestinal mucosal epithelial cells and endothelial cells are the structural basis of intestinal mucosal barrier function. The connections between cells include tight junctions, gap junctions, adherens junctions, and desmosomes, among which tight junctions are the most important connection between cells. The disorder of tight junction function and structure is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L33/135A23L2/38A23C9/123A23L11/65A23L11/50A23C19/00A61K35/747A61P1/00C12R1/225
CPCC12N1/20A23L33/135A23L2/382A23C9/1234A23C11/106A23C19/00A61K35/747A61P1/00A23V2002/00C12R2001/225C12N1/205A23V2400/143A23V2200/3204
Inventor 翟齐啸陈卫赵岩于雷雷陆文伟田丰伟赵建新张灏
Owner JIANGNAN UNIV
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