Lactobacillus plantarum with abilities of reducing cholesterol and promoting intestinal short chain fatty acid generation ability and application of lactobacillus plantarum

A technology of Lactobacillus plantarum and short-chain fatty acids, applied in the directions of Lactobacillus, application, microorganism, etc., can solve the problem of low incidence of cardiovascular disease, and achieve the goal of reducing serum cholesterol level, high cholesterol-lowering ability, and relieving liver pathological conditions. Effect

Active Publication Date: 2018-11-02
SICHUAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Herdsmen in Daocheng, Sichuan have long eaten homemade cheese with high fat content (butter milk cake, which is fermented from unrefined yak milk without adding exogenous lactic acid bacteria), but the incidence of cardiovascular disease is not high

Method used

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  • Lactobacillus plantarum with abilities of reducing cholesterol and promoting intestinal short chain fatty acid generation ability and application of lactobacillus plantarum
  • Lactobacillus plantarum with abilities of reducing cholesterol and promoting intestinal short chain fatty acid generation ability and application of lactobacillus plantarum
  • Lactobacillus plantarum with abilities of reducing cholesterol and promoting intestinal short chain fatty acid generation ability and application of lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation, Screening and Molecular Biological Identification of Lactobacillus plantarum CGMCC 15463:

[0028] 1. Material preparation

[0029] Milk cake samples were provided by Kalong Village, Mula Township, Daocheng County, Sichuan Province;

[0030] The universal primer pair was synthesized by Invitrogen (Shanghai), and the 27F / 1492R sequence is as follows:

[0031] 27F: AGAGTTTGATCMTGGCTCAG

[0032] 1492R: TACGGYTACCTTGTTACGACTT

[0033] MRS liquid medium: glucose 20.0 g, peptone 10.0 g, beef extract 10.0 g, yeast powder 6.0 g, Tween 801.0 mL, dipotassium hydrogen phosphate 2.0 g, ammonium citrate 2.0 g, sodium acetate 5.0 g, magnesium sulfate 0.58 g , manganese sulfate 0.25 g, pure water 1 L, pH natural (add 1.5%-2.0% agar for solid medium).

[0034] 2. Specific ways

[0035]Weigh 10.0 g of the traditional yak milk cheese (milk cake) sample, stir it thoroughly in a conical flask filled with 90 mL of sterile water, and then shake it on a shaker for 60 min to co...

Embodiment 2

[0039] Detection of main fermentation characteristics of Lactobacillus plantarum CGMCC 15463

[0040] PBS buffer solution: Potassium dihydrogen phosphate 0.20 g, disodium hydrogen phosphate 1.15 g, sodium chloride 8.0 g, potassium chloride 0.2 g, add 800 mL of pure water and stir to dissolve, adjust concentrated hydrochloric acid to the corresponding pH value, and dilute to 1 L.

[0041] DMEM complete medium: DMEM stock solution, 10% fetal bovine serum, 1% 100 U / ml double antibody.

[0042] 1. Acid resistance test: Inoculate the activated third-generation lactic acid bacteria into PBS buffer solution with pH 3.5, 2.5, and 2.0 at an inoculum amount of 1%, respectively, culture at 37°C for 3 hours, and measure the number of viable lactic acid bacteria by gradient dilution plate method.

[0043] 2. Bile salt resistance test: Inoculate the activated three-generation lactic acid bacteria into the MRS liquid medium containing 0.1%, 0.2%, and 0.3% bovine bile salt at an inoculation ...

Embodiment 3

[0053] Determination of Cholesterol Degradation Ability of Lactobacillus plantarum CGMCC 15463 in Vitro

[0054] MRS-cholesterol medium: MRS liquid medium (same as Example 2) was added with 0.1% ox bile salt, and cholesterol was added to make the final concentration 100 μg / mL.

[0055] The third-generation activated lactic acid bacteria were inoculated into MRS-cholesterol medium at an inoculum of 1% by the o-phthalaldehyde method, and cultured on a shaker at 120 r / min at 37°C for 24 h. Centrifuge at 10,000 g for 10 min at 4°C, take 1 mL of the supernatant into a stoppered colorimetric tube, add 1 mL of 33% KOH and 2 mL of absolute ethanol, vortex for 90 s, place in a water bath at 37°C for 15 min, and cool to At room temperature, add 2 mL of sterile water and 3 mL of n-hexane, vortex for 60 s, let stand for 10 min, take 1 mL of the upper layer liquid in a new test tube and evaporate to dryness in a water bath at 65°C, add 2 mL of o-phthalaldehyde-glacial acetic acid solution ...

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Abstract

The invention relates to the technical field of microorganisms, and particularly discloses lactobacillus plantarum N-1 with abilities of reducing cholesterol and promoting intestinal short chain fattyacid generation ability and application of the lactobacillus plantarum. The lactobacillus plantarum N-1 was deposited in the China General Microbiological Culture Collection Center UNDER the accession number of CGMCC NO. 15463 with a deposit date of 20 Mar. 2018. The strain is separated from a traditional yak milk cheese product of Kalong Village, Mula Township, Daocheng County, Garze Tibetan Autonomous Prefecture, Sichuan Province, grows well on an MRS agar medium, has certain tolerance capacity on acid and bile salt, has high adhesion ability to human colon cancer cells Caco-2, has high cholesterol reducing ability, can remarkably improve generation of intestinal short chain fatty acids, is applied to the field of functional food, not only has actual production value, but also has quiteimportant significance on health of human bodies.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Lactobacillus plantarum capable of lowering cholesterol and promoting the production of intestinal short-chain fatty acids, and in particular to the field of production and development of functional lactic acid bacteria and their products. Background technique [0002] Cardiovascular disease is the number one killer of human health and life, and hypercholesterolemia is one of the main risk factors for cardiovascular disease. With the Westernization of diet, the prevalence of dyslipidemia in my country is rapidly increasing, gradually approaching the level of Western countries, seriously endangering human health. [0003] Lactic acid bacteria (LAB) is a general term for a class of spore-free, Gram-positive bacteria that can use fermentable carbohydrates to produce large amounts of lactic acid. As a type of probiotics, lactic acid bacteria are recognized as high...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L33/135C12R1/25
CPCA23L33/135A23V2002/00C12R2001/25C12N1/205A23V2400/169A23V2200/3262A23V2200/32
Inventor 孙群刘绒梅田圆圆张国建梁鹏宽周智威
Owner SICHUAN UNIV
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