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Method for directionally and rapidly screening lactobacillus plantarum capable of producing bile salt hydrolase

A technology of Lactobacillus plantarum and bile salt hydrolysis enzyme, applied in the field of microorganisms, can solve the problems of missing lactic acid bacteria, heavy separation workload, and lactic acid bacteria may not have the ability to lower cholesterol, etc., to achieve the effect of small workload and reduce blindness

Inactive Publication Date: 2018-06-12
HENAN UNIV OF SCI & TECH
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Problems solved by technology

Through comparison, it is found that the operation of method 1) is relatively cumbersome, and it is necessary to test the cholesterol-lowering ability of each bacterial strain isolated and purified, but there will be no omissions. Although the latter two methods are easy to operate, they must be tested first. Separation and purification of lactic acid bacteria, and then screening of lactic acid bacteria with cholesterol-lowering ability by indicator method or calcium precipitation method, the initial isolation workload is relatively large, and the isolated lactic acid bacteria may not have cholesterol-lowering ability, so most of the previous work is useless , and at the same time, the target strain or even the lactic acid bacteria with strong degradation effect may be missed
Therefore, none of the above three methods can be called drow effective

Method used

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  • Method for directionally and rapidly screening lactobacillus plantarum capable of producing bile salt hydrolase
  • Method for directionally and rapidly screening lactobacillus plantarum capable of producing bile salt hydrolase
  • Method for directionally and rapidly screening lactobacillus plantarum capable of producing bile salt hydrolase

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Embodiment 1

[0039] A method for directional and rapid screening of Lactobacillus plantarum producing bile salt hydrolysis enzyme, comprising the following steps:

[0040] The intestinal contents of different animals (i.e. fresh feces) were used as separate samples:

[0041] 1) Collection of total microorganisms in samples and acquisition of metagenomics

[0042] Aseptically take about 3 grams of fresh feces from different animals, grind them aseptically, place them in 50mL centrifuge tubes, add 7mL PBS solution, vortex fully for 5min, centrifuge at 500r / min for 3min, take the supernatant, repeat the above steps once The supernatant was centrifuged at 6000r / min for 5min, the supernatant was discarded, the precipitated thalline was transferred to a clean centrifuge tube, and the total microbial metagenomic DNA of the intestinal contents of the animal was extracted by thermal cracking (see Molecular Cloning Experiment Guide, J. Sambrook et al.);

[0043] 2) Confirmation of BSH-containing L...

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Abstract

The invention discloses a method for directionally and rapidly screening lactobacillus plantarum capable of producing bile salt hydrolase, and belongs to the technical field of microorganisms. The invention comprises the following steps: (1) taking metagenome DNA of a sample as a template, performing PCR amplification on BSHL / BSHR by utilizing a primer, and screening to obtain the sample with a BSH gene; (2) taking the sample as a separating object, and purifying to obtain a suspected plant lactobacillus strain; (3) taking the strain genome DNA as a template, performing PCR amplification on the BSHL / BSHR by utilizing the primer, and screening to obtain a suspected strain with the BSH gene; (4) performing molecular identification on the strain and screening to obtain the lactobacillus plantarum capable of producing bile salt hydrolase. According to the method, most samples which do not contain the lactobacillus plantarum capable of producing the BSH can be filtered out through the firststep of amplification of metagenome-specific fragments, so that blind screening is avoided, the screening range is greatly reduced, and the method has the advantages of directionality, rapidness, easiness, convenience, low workload and the like.

Description

technical field [0001] The invention relates to a method for directional and rapid screening of lactobacillus plantarum producing bile salt hydrolyzing enzyme, which belongs to the technical field of microorganisms. Background technique [0002] In recent decades, cardiovascular disease has become the leading cause of death for all human beings. Clinical trials have shown that high serum cholesterol levels are the main pathogenic factors causing cardiovascular diseases. When the body exceeds the normal value of serum cholesterol level (0-5.2mmo / L), it will show high cholesterol, which will cause the body to suffer from kidney disease, heart disease, stroke and so on. For every 1% increase in cholesterol levels in the body, the risk of cardiovascular disease will increase by 35%. The survey found that in 2012, the number of deaths caused by cardiovascular diseases worldwide (about 17.5 million) accounted for about 31% of the total number of human deaths, of which nearly 42%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/25
CPCC12N1/20
Inventor 丁轲陈亚茹刘炜余祖华李旺李元晓何万领曹平华赵龙妹王玉琴张春杰程相朝
Owner HENAN UNIV OF SCI & TECH
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