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A kind of Lactobacillus johnsonii and application thereof

A technology of Lactobacillus johnsonii and Lactobacillus johnsonii, applied in the field of microbiology, can solve problems such as increased platelet aggregation, high choline level and increased betaine level

Active Publication Date: 2022-06-24
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, choline supplementation can lead to increased platelet aggregation, so this risk is currently suppressed by taking aspirin
Reduction of high blood choline levels and concomitant increase in betaine levels caused by a high-choline diet by probiotics has not been reported

Method used

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  • A kind of Lactobacillus johnsonii and application thereof
  • A kind of Lactobacillus johnsonii and application thereof
  • A kind of Lactobacillus johnsonii and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Isolation and identification of Lactobacillus in the intestinal tract of C57B6 / J mice

[0045] Fresh C57B6 / J mouse intestinal contents were diluted with sterile water at room temperature. Under sterile conditions, 100 mg of mouse intestinal contents were taken into test tubes. 1 mL of sterile water was added, and the sample was centrifuged at 4000 rpm for 1 minute. Each 50 μL of the supernatant was spread on MRS plate and cultured at 37°C for 24 hours. By observing the characteristics of the colonies on the plate, the colonies were screened from the plate for molecular biological identification.

[0046] The genomic DNA of EU03 strain was extracted by ctab / sds method and used as a template for PCR amplification. The 16s ribosomal DNA sequence of the strain was amplified and sequenced using universal primers 27f: 5'-agagttttgatggctcag-3' (SEQ ID NO: 1) and 1492r: 5'-acggttatccttgttaccgatt-3' (SEQ ID NO: 2). Amplification was performed using polymerase chain...

Embodiment 2

[0048] Example 2: Modulating effect of Lactobacillus johnsonii EU03 on intestinal flora diversity in healthy and MI rats

[0049] The experimental animals were 6-week-old male healthy SD rats with a body weight of 240 ± 10 g, provided by Beijing Weitong Lihua Animal Experiment Center. The rats were reared in the Experimental Animal Center of Tianjin University of Traditional Chinese Medicine (Tianjin) at room temperature (22±2°C), relative humidity of 58-65%, and photoperiod of 12h alternating. In this study, 3-6 identically treated rats were tested in each group.

[0050] Preparation of Lactobacillus agent: Lactobacillus johnsonii EU03 was inoculated into MRS liquid medium according to 2% inoculum, cultured at 37°C for 24h, taken out and centrifuged (10000r / min, 10min, 4°C) to discard the supernatant, After washing twice with sterilized water, adjust the bacterial concentration to 10 with sterilized water. 8 CFU / mL (colony count: transfer the cryopreserved Lactobacillus joh...

Embodiment 3

[0060] Example 3: Modulating effect of Lactobacillus johnsonii EU03 on the structure of intestinal flora in healthy and MI rats

[0061] Use Uparse software (Uparse v7.0.1001) to cluster all the Effective Tags of all samples. By default, the sequences are clustered into OTUs with 97% identity. At the same time, the representative sequences of OTUs will be selected, according to the algorithm principle. , and screen the sequence with the highest frequency in OTUs as the representative sequence of OTUs. Species annotation analysis was performed on the representative sequences of OTUs using the Mothur method and the SSUrRNA database of SILVA (http: / / www.arb-silva.de / ) (with a threshold of 0.8 to 1), and taxonomic information was obtained and analyzed at each taxonomic level. : phylum (door) genus (genus) statistics on the community composition of each sample.

[0062] Experimental results:

[0063] image 3 A shows the most abundant phyla in the rat gut microbiota, accounting ...

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Abstract

The invention belongs to the technical field of microorganisms, and discloses a Lactobacillus johnsonii EU03 and an application thereof. Lactobacillus johnsonii described in the present invention can improve Lactobacillus (Lactobacillus), Muribaculaceae (p<0.05), Akkermansia (Akkermansia), Allobaculum, Bacteroides (Bacteroides) (p<0.05) and Blautia in intestinal flora Equal the abundance of dominant intestinal flora, reduce the abundance of Romboutsia, Lachnospiraceae_NK4A136 and other flora, have a positive regulatory effect on the structure of intestinal flora, thereby improving symptoms such as metabolic syndrome; and affect the intestinal flora related metabolites It can reduce the concentration of trimethylamine oxide (TMAO)-related metabolite choline, increase the concentration of betaine, increase the concentration of short-chain fatty acids (SCFAs) in the intestinal flora, and play a role in alleviating cardiovascular diseases.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, in particular to a Lactobacillus johnsonii EU03 and applications thereof, in particular to a Lactobacillus johnsonii capable of regulating intestinal flora and its metabolites and applications thereof. Background technique [0002] The intestinal flora is closely related to many diseases, mainly including gastrointestinal diseases, immune diseases, osteoporosis, cardiovascular diseases, metabolic diseases, malignant tumors and neurological diseases. Intestinal flora affects the host's microbial ecosystem mainly by enhancing immune function, preventing or treating intestinal diseases such as diarrhea, constipation, and flatulence, helping nutrient absorption, and regulating metabolic abnormalities caused by intestinal flora imbalance such as bone. metabolism and glucose and lipid metabolism. Among the intestinal probiotics, Lactobacillus sp. is a Gram-positive bacteria and is classified as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/747A61P1/00A61P9/00A61P31/04A61P3/00A23C9/123A23L11/50A23L19/00A23L33/135C12R1/225
CPCA61K35/747A61P1/00A61P9/00A61P31/04A61P3/00A23C9/1234A23L19/00A23L33/135A23V2002/00A23V2400/151A23V2200/30A23V2200/328A23V2200/332A23V2200/3204
Inventor 赵鑫王小莹高秀梅钟鑫勤魏兴
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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