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Lactobacillus reuteri and application thereof

A technology of Lactobacillus reuteri and yogurt, applied in the field of Lactobacillus reuteri and its application, can solve the problems of hearing harm, human flora imbalance, antibiotic abuse, etc., and achieve excellent bacteriostatic effect and excellent bacteriostatic effect. Effect

Active Publication Date: 2019-10-25
深圳华大基因农业控股有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The abuse of antibiotics is a serious problem facing the world at present. Experts point out that the abuse of antibiotics will not only cause drug resistance, but also cause the imbalance of human flora, destroy the micro-ecological balance, and cause harm to human hearing, liver, kidney, etc.

Method used

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  • Lactobacillus reuteri and application thereof
  • Lactobacillus reuteri and application thereof
  • Lactobacillus reuteri and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Embodiment 1: the source and identification of bacterial strain

[0051] Healthy adult feces samples were taken as the research object, and suspected colonies of lactic acid bacteria were isolated and screened, and then 16SrDNA amplification was used to identify the suspected strains picked. The details are as follows:

[0052] (1) Medium formula

[0053] The inventors prepared optimized MRS solid medium and MRS culture fluid.

[0054] MRS solid medium, referred to as MRS medium, the formula for 1L is: casein peptone 10.0g / L, beef extract 10.0g / L, yeast extract 5.0g / L, glucose 20.0g / L, dipotassium hydrogen phosphate 2.0g / L, Tween 80 1.0g / L, triammonium citrate 2.0g / L, sodium acetate 5.0g / L, magnesium sulfate 0.1g / L, manganese sulfate 0.05g / L, agar 17.5g.

[0055] MRS broth medium, that is, the formula of MRS culture medium is only that agar is not added to MRS medium, and the rest are the same.

[0056] The prepared medium and culture solution were adjusted to pH 6...

example 16

[0059] The primer sequences for 16s rDNA amplification in this example are shown in SEQ ID NO: 1 and SEQ ID NO: 2.

[0060] 5'-AGAGTTTGATCATGGCTCAG-3' (SEQ ID NO: 1).

[0061] 5'-TAGGGTTACCTTGTTACGACTT-3' (SEQ ID NO: 2).

[0062] Primers were synthesized by Shenzhen BGI Institute.

[0063] The PCR reaction system is as follows: 50 μL reaction solution includes: 1 μL of 10 mM dNTPs, 5 μL of 10×buffer, 1 μL of 10 mM upstream and downstream primers, 1 μL of bacteria solution PCR template, 1 μL of 5 U / μL Taq enzyme, supplemented with ddH 2 O supplemented to 50 μL.

[0064] The PCR reaction conditions are: pre-denaturation at 94°C for 5 minutes, and then enter 35 cycles: 94°C for 30s, 60°C for 30s, and 72°C for 1min. After the cycle, extend at 72°C for 5min, and stand by at 4°C.

[0065] The PCR amplification products were recovered by gel cutting, and the recovered PCR products were sequenced. In this example, the kit TaKaRa MiniBEST agarose GeL DNA Extraction kit was used for...

Embodiment 2

[0085] Embodiment 2: in vitro acid resistance, bile salt resistance experiment

[0086] The inventors evaluated the in vitro acid resistance and bile salt tolerance of the Lactobacillus reuteri WX-94 isolated in Example 1, specifically as follows:

[0087] Preparation of simulated gastric juice SGF: In 1000mL simulated gastric juice, 3.2g of pepsin, 2.0g of NaCl, and HCl were adjusted to pH 3.0 and pH 2.0, and then filtered through a 0.2μm microporous membrane to obtain the SGF of this example liquid, set aside.

[0088] Pick the slant strain of Lactobacillus reuteri WX-94 in the MRS culture medium and culture at 37°C for 16-18 hours. Centrifuge the bacterial suspension at 4000r / min for 15min, remove the supernatant, weigh the wet weight of the bacterial cell, resuspend the bacterial cell in normal saline at a ratio of 0.1g / mL, and then add it to the SGF solution at a ratio of 1:10, fully After mixing, they were cultured in a constant temperature incubator at 37°C. Cells wer...

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Abstract

The invention provides lactobacillus reuteri. The lactobacillus reuteri was deposited at the General Microbiology Center of China Microbiological Culture Collection and Management Committee on December 14, 2017 with the deposition number of CGMCC No. 15062. The lactobacillus reuteri has broad-spectrum bacteriostatic effect, good adhesion property and strong acid resistance and bile salt resistance, is obviously superior to an existing commercial strain, and is a newly isolated and discovered active bacterium with a good probiotic function.

Description

technical field [0001] The present invention relates to the field of microorganisms, specifically, the present invention relates to Lactobacillus reuteri and its application, more specifically, the present invention relates to Lactobacillus reuteri, the application and medicine of Lactobacillus reuteri in inhibiting pathogenic bacteria or health care , Lactobacillus reuteri tablets, compound probiotic tablets, food, yogurt, health products, food additives, compositions, and Lactobacillus reuteri in the preparation of medicines, foods, health products, food additives or compositions use. Background technique [0002] The misuse of antibiotics is a serious problem facing the world at present. Experts point out that the misuse of antibiotics will not only cause drug resistance, but also cause the imbalance of human flora, destroy the micro-ecological balance, and cause harm to human hearing, liver, and kidneys. Therefore, reducing the use of antibiotics has become a world tren...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P31/04A61P31/10A23L33/135A23L3/3571A23C9/123C12R1/225
CPCA61P31/04A61P31/10A23L33/135A23L3/3571A23C9/123A61K2035/115C12R2001/225C12N1/205A23V2400/173Y02A50/30
Inventor 苏敦彭灿周小兵何小芬关卫霞徐敏肖晨李卉董扬
Owner 深圳华大基因农业控股有限公司
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