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Lactobacillus plantarum ZJUFYJ7 and application thereof

A technology of Lactobacillus plantarum and drugs, applied in the fields of bacteria, microbes, food science, etc., can solve the problems of postprandial blood sugar level reduction, glucose absorption delay, genome and functional differences, etc., to relieve hyperglycemia and insulin resistance, improve intestinal Effects of intestinal flora disturbance

Pending Publication Date: 2022-04-12
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that α-amylase and α-glucosidase inhibitors can slow the release of glucose from starch and oligosaccharides, resulting in delayed glucose absorption and lower postprandial blood glucose levels
[0006] However, there are great differences in the genome and function of different strains of Lactobacillus plantarum, and no Lactobacillus plantarum that can alleviate insulin resistance has been reported yet

Method used

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  • Lactobacillus plantarum ZJUFYJ7 and application thereof
  • Lactobacillus plantarum ZJUFYJ7 and application thereof
  • Lactobacillus plantarum ZJUFYJ7 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Isolation and Identification of Bacterial Strains

[0047] 1. Screening and isolation of strains

[0048] Material: Kimchi, a traditional fermented food commonly eaten in the south, the present invention uses samples from Sichuan.

[0049] Weigh 5g kimchi and add it to 45mL 0.85% sterilized physiological saline, vibrate to make a suspension, carry out serial gradient dilution, take 10% after dilution -5 、10 -6 、10 -7 Three gradients were spread on the MRS medium, and each gradient was made in triplicate. Put the coated plate into an anaerobic incubator at 37°C for 24-48h. The colonies with obvious differences were selected and separated by streaking on the MRS medium, and the plate was placed in an anaerobic incubator at 37°C for 24-48h. After streaking for 3-5 times in a row, take a drop of 5% hydrogen peroxide that is now prepared and drop it on the glass slide, pick a single colony on the solid medium and insert it into the 5% hydrogen peroxide drop an...

Embodiment 2

[0060] Example 2 Tolerance of simulated artificial gastrointestinal fluid

[0061] The artificial simulated gastrointestinal juice needs to be freshly prepared and must pass through a membrane (0.22μm) to sterilize.

[0062] Simulated gastric juice (GJ): Pepsin (1:10000) was added to PBS (pH2.5) at a concentration of 3.5g / L.

[0063] Simulated intestinal juice (IJ): NaHCO 3 11g / L, NaCl 2g / L, trypsin 1g / L, pig bile salt 18g / L, adjust the pH value to 8.0, and sterilize by 0.22μm membrane filtration for later use.

[0064] After refrigerated centrifugation, live cells were obtained, washed twice with PBS (pH7.4), and the concentration of live cells was adjusted to about 10 9 CFU / mL, add 0.5mL bacterial suspension to 4.5mL simulated gastric juice, and place in an anaerobic incubator for cultivation at 37°C. At 0, 1.5h, and 3h respectively, the MRS agar pour plate method was used to count.

[0065] After GJ treatment for 3 hours, take 0.5 mL of GJ culture solution and add it t...

Embodiment 3

[0071] Example 3 Determination of Adhesion Ability

[0072] Caco-2 cells were grown in IMDM medium containing 15% fetal bovine serum and 1% double antibody at 37°C and 5% CO 2 Cultured in an incubator with 0.25% trypsin containing 0.02% EDTA and passaged at a ratio of 1:3 when the cell proliferation and fusion rate reached about 80%. Cells in the logarithmic growth phase were used for the experiment.

[0073] Caco-2 cells at 2.5×10 5 cells / well seeded in 24-well plate, change medium every day, 37°C 5% CO 2 Cultured for 5 days to reach a monolayer of cells. After the strain was cultured overnight, centrifuge at 4°C and 8000 rpm for 10 min, wash the bacteria twice with sterile PBS, and adjust the bacterial concentration to about 1×10 with IMDM cell culture medium (without double antibody). 8CFU / mL and counted on the agar plate. Wash the plate 5 times with PBS to remove the residual double antibody on the plate, then add 1 mL of the prepared cell culture suspension containin...

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Abstract

The invention discloses a lactobacillus plantarum ZJUFYJ7 and an application of the lactobacillus plantarum ZJUFYJ7, and belongs to the technical field of biology. The preservation number of the lactobacillus plantarum ZJUFYJ7 is CCTCC (China Center for Type Culture Collection) NO: M 2020127. The lactobacillus plantarum ZJUFYJ7 provided by the invention has a very high survival rate and relatively strong gastrointestinal adhesive power in a gastrointestinal tract environment, can remarkably inhibit the reproduction of gastrointestinal pathogenic bacteria, has relatively strong free radical scavenging ability, improves the sugar intake of HepG2 hepatocytes and promotes the secretion of IL-10 by macrophages, and has a potential anti-inflammatory effect. Animal experiments prove that the lactobacillus plantarum ZJUFYJ7 can remarkably relieve hyperglycemia and insulin resistance induced by high-fat diet, and through colonization of the lactobacillus plantarum, intestinal flora disorder is improved, and bile acid and short-chain fatty acid metabolism is regulated and controlled, so that the effect of reducing blood sugar is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to plant lactobacillus (Lactobacillus plantarum) ZJUFYJ7 and application thereof. Background technique [0002] With the change of people's dietary structure, the intake of high-fat diet has increased significantly, and the resulting insulin resistance has been increasing year by year. Type 2 diabetes is a chronic metabolic disorder characterized clinically by hyperglycemia often accompanied by low-grade inflammation. Studies have shown that metabolic disorders of the body are closely related to the decline in the ability of immune cells to produce interleukin-10 (IL-10) (Verma et al., 2016). Inhibition of IL-10 will promote the expression of inflammatory factors such as TNF-α and IL-6, and the insulin signal will be deteriorated, and even activate the gluconeogenesis and lipogenesis pathways (Gotoh et al., 2017). [0003] In recent years, there have been more and more studies on the ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A23L33/135A61P31/04A61P1/00A61P29/00A61P3/10A61P3/00C12R1/25
CPCY02A50/30
Inventor 冯凤琴钟浩张峻珲赵敏洁刘滔沈飞王倩倩
Owner ZHEJIANG UNIV
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