Method for fast screening lactobacillus for degrading cholesterol

A technology for lowering cholesterol and lactic acid bacteria, which is applied in the direction of microbial-based methods, biochemical equipment and methods, and microbial measurement/inspection. small effect

Inactive Publication Date: 2010-03-17
DALIAN POLYTECHNIC UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, traditional screening techniques were used for the screening of cholesterol-lowering lactic acid bacteria. The screening cycle was long and the workload was heavy. Breeding high-performance lactic acid bacteria strains was like finding a needle in a haystack.

Method used

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  • Method for fast screening lactobacillus for degrading cholesterol
  • Method for fast screening lactobacillus for degrading cholesterol
  • Method for fast screening lactobacillus for degrading cholesterol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Using fermented milk products such as probiotic yogurt and cheese as raw materials for separation

[0033] (1) Enrichment culture of lactic acid bacteria:

[0034] Weigh 10-20 grams of yogurt, cheese and other foods rich in lactic acid bacteria as enrichment samples, add them to 100-200ml skim milk or MRS liquid medium for enrichment culture, and cultivate them at 37°C for 48 hours before using them as isolated samples.

[0035] Skimmed milk medium: add 10 grams of skimmed milk powder to 100ml of distilled water, and sterilize at 112°C for 20 minutes.

[0036] MRS liquid medium: peptone 10g, beef extract 10g, yeast extract 5.0g, glucose 20.0g, K 2 HPO 4 2.0g, NaOAC 5.0g, MgSO 4 0.2g, Tween 80 1.0g, triammonium citrate 2.0g, distilled water 1000mL.

[0037] (2) Preliminary isolation and purification of cholesterol-lowering lactic acid bacteria:

[0038] Aseptically take 0.5 or 1ml of the above-mentioned enriched culture solution, add it to a test tube co...

Embodiment 2

[0063] Embodiment 2 takes pickles, spicy cabbage and other fermented vegetables as separation raw materials

[0064] (1) Enrichment culture of lactic acid bacteria:

[0065] Measure 10-20ml of fermented vegetable juices such as kimchi juice and spicy cabbage juice, dilute with sterile distilled water, draw 2ml into 20ml of sterilized MRS liquid medium for enrichment culture by aseptic operation, and let it stand at 37°C Cultured for 48h, used as an isolated sample.

[0066] (2) Preliminary isolation and purification of cholesterol-lowering lactic acid bacteria:

[0067] The specific steps are the same as Example 1

[0068] (3) Colony PCR rapid screening of bile salt hydrolase (BSH) gene

[0069] ①BSH gene-specific primer design

[0070] The specific steps are the same as Example 1

[0071] ②Strain PCR amplification

[0072] Select the isolated and purified lactic acid bacteria, pick a single colony with a sterile toothpick into a 0.2mL PCR tube, add 10 μL of sterile 1×TE...

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Abstract

The invention discloses a method for fast screening lactobacillus for degrading cholesterol, comprising the steps of: preliminarily separating purified lactobacillus; selecting single bacterial colonyinto 0.2 mL PCR tube; adding with 10 mu L sterile 1*TE buffer solution and evenly blending; boiling at 100 DEG C for 5 min; cooling at -20 DEG C for 5 min; circling for 2-3 times; decentralizing at 5000r / min for 1 min; absorbing 8uL of supernatant to be taken as DNA template of PCR amplification; executing the PCR amplification; and analyzing and confirming the positive and negative of the BSH gene by means of electrophoresis experiment. The method further confirms the bacterial strain capable of degrading the cholesterol and can confirm the attribute of lactobacillus bile salt hydrolase in 12 hours. The method has prominent advantages of short work period and less workload.

Description

technical field [0001] The invention relates to a rapid screening method for microorganisms, in particular to a rapid screening method for cholesterol-lowering lactic acid bacteria. Background technique [0002] According to the 1996 Health Statistical Yearbook published by the World Health Organization, 15.3 million people died of cardiovascular diseases worldwide, accounting for 29% of the total deaths. Therefore, cardiovascular diseases rank first among the top five causes of death in the world, and their common pathological basis is arteriosclerosis. Atherosclerosis is caused by many factors, among which the most important risk factor is hyperlipidemia, that is, the increase of blood cholesterol and blood triglycerides. Therefore, keeping the cholesterol concentration in the blood within the normal range is crucial for preventing atherosclerosis and cardiovascular and cerebrovascular diseases. [0003] Nutrition and medical experts in the United States and my country r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/225
Inventor 侯红漫郭东启张公亮
Owner DALIAN POLYTECHNIC UNIVERSITY
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