Lactobacillus plantarum RS-09 with high adhesion and applications thereof

A technology of Lactobacillus plantarum and Lactobacillus plantarum, applied in the field of microorganisms, can solve problems such as side effects and drug resistance enhancement, and achieve the effects of strong acid production ability, enhanced resistance and strong adhesion ability

Active Publication Date: 2019-05-03
SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, antibiotic intervention is the routine clinical treatment of salmonellosis, but the use of antibiotics will be accompanied by serious side effects, such as increased drug resistance

Method used

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  • Lactobacillus plantarum RS-09 with high adhesion and applications thereof
  • Lactobacillus plantarum RS-09 with high adhesion and applications thereof
  • Lactobacillus plantarum RS-09 with high adhesion and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Lactobacillus plantarum ( Lactobacillus plantarum ) Isolation, purification and identification of RS-09

[0043] Isolation and purification of Lactobacillus plantarum strains: Mix the apple juice fermentation broth, take 5 mL of the fermentation broth, and serially dilute it 10 times to 10 -6 , take 200 μL of each dilution concentration and spread it on the solution containing 1% CaCO 3 Lactic acid bacteria selective medium MRS medium, cultivated in 37°C anaerobic tank for 48 hours, picked a single colony on the plate, streaked on MRS solid medium, cultured in 37°C anaerobic tank for 48 hours, and obtained pure colonies; The purified colonies were identified by Gram staining; the strains contained CaCO3 The diameter of the colony on the MRS medium is 1-2mm, white, raised, and has a calcium-dissolving circle; the strain is Gram-positive, and the round end is straight rod-shaped; the Gram staining characteristics of the bacteria are as follows: figure 1 shown. Scannin...

Embodiment 2

[0049] Lactobacillus plantarum ( Lactobacillus plantarum ) RS-09 Adhesion Ability Determination

[0050] Take a sterile 6-well plate, place a sterile cover glass in each well, and passage once at a concentration of 1×10 5 cells / mL of the human colon cancer cell line HT-29 was inoculated into a 6-well plate in 5% CO 2 Cultivate overnight at 37°C to a monolayer of cells, discard the culture medium, wash gently with DMEM solution three times, add 200 μL of Lactobacillus plantarum suspension (5 ×10 7 CFU / well) and 1800 μL DMEM culture solution, repeated for three wells, placed at 37°C, 5% CO 2 After culturing in the incubator for 2 hours, discard the supernatant, wash with sterile PBS three times, add 400 μL of 0.25% trypsin to each well to digest for 5 minutes, and then add 400 μL of DMEM solution containing 10% fetal bovine serum to terminate the digestion; take cell slides, Carry out Gram staining, randomly select 20 fields of view under the microscope, count the total num...

Embodiment 3

[0052] Lactobacillus plantarum ( Lactobacillus plantarum ) Dynamic distribution of RS-09 intestinal colonization

[0053] cFDA-SE labeled Lactobacillus plantarum RS-09: take 2×10 8 CFU / mL RS-09 bacterial solution 200ml, centrifuge at 4000rpm for 10min at 4°C, wash twice with cold PBS (pH7.2), resuspend in 20mL cFDA-SE solution with a final concentration of 20μmol / L, Incubate at 37°C in the dark for 20 min, wash with PBS three times to remove unbound cFDA-SE molecules, resuspend in PBS to a final concentration of 2×10 9 CFU / mL.

[0054] Dynamic distribution of colonization in different parts of the intestinal tract of Lactobacillus plantarum RS-09 mice: 54 BALB / c mice with SPF grade at 4 to 5 weeks old were selected and fed with cFDA-SE-labeled Lactobacillus plantarum suspension (2× 10 9 CFU / mL), 500 μL for each mouse; the intestines of the mice were taken at 5 different time points (2h / 12h / 24h / 48h / 96h) after gavage, 3 mice at each time point; The intestinal segments of th...

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Abstract

The invention relates to a lactobacillus plantarum RS-09 with high adhesion and applications thereof, and belongs to the field of microorganisms. The strain is separated from apple juice, and is preserved in in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO.17118; has stronger acid resistance and choline salt resistance, can be stored for a longtime, has higher adhesion capacity to human colon cancer cells HT-29, and can be stably planted in intestinal tracts, wherein the planting rate of the cecum is highest; can obviously reduce the content of TNF-alpha in serum, increase the content of IL-10 and IL-4 in the serum, and has the function of immunoregulation; and has strong bacteriostatic effect on salmonella enteritis, has prevention andtreatment effects on enteritis diseases caused by flora disorder, can be applied to the field of functional foods, not only has practical production value, but also has very important significance onbody health.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a strain of Lactobacillus plantarum ( Lactobacillus plantarum ) and its applications. Background technique [0002] Lactic acid bacteria are probiotics in the gastrointestinal tract of animals. They can metabolize organic acids, antibacterial peptides, diacetyl, bacteriocins and other natural antibacterial active substances. They have health care functions such as maintaining the balance of normal intestinal flora and enhancing immunity. With the improvement of people's health awareness, the requirements for healthy food are gradually increasing. As the main force of functional food and medicine, lactic acid bacteria are considered as a natural feed additive and food additive with broad application prospects. [0003] Salmonella, belonging to the Enterobacteriaceae family, is a common zoonotic pathogen that causes intestinal diseases. In China, 22.2% of food poisoning is caused b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02A23L33/135A23K10/18A61K35/747A61P31/04A61P1/00A61P37/04C12R1/25
Inventor 姜琳琳张兴晓赵小雨陈国忠张建龙于馨朱洪伟黄清荣
Owner SHANDONG YANGCHENG BIOLOGY TECH CO LTD
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