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Lactobacillus plantarum capable of antagonizing campylobacter jejuni and inhibiting expression of flaA gene of campylobacter jejuni

A technology of Campylobacter jejuni and Lactobacillus plantarum, applied in the field of microorganisms, can solve the problems of not involving Campylobacter jejuni, not many, reducing Campylobacter jejuni antigens, etc., and achieve good adhesion characteristics, growth inhibition, and wide application prospects

Active Publication Date: 2016-01-13
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, some published patent documents relate to the antagonism of Campylobacter jejuni and its use. For example, CN104870472A discloses an antibody and its fragments specific to Campylobacter jejuni; CN102497875A discloses an antigen for reducing the colonization of Campylobacter jejuni; CN103381220A discloses A kind of medicine that can treat Campylobacter jejuni of chicken; However, there are not many patent application documents related to lactic acid bacteria antagonizing Campylobacter jejuni and inhibiting the expression of its virulence gene. CN102533618A discloses a plant with the ability to inhibit Campylobacter jejuni in vitro Lactobacillus, but the patent does not involve the expression of the flaA gene of Campylobacter jejuni and its efficacy in cell models and in vivo antagonism against Campylobacter jejuni

Method used

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  • Lactobacillus plantarum capable of antagonizing campylobacter jejuni and inhibiting expression of flaA gene of campylobacter jejuni
  • Lactobacillus plantarum capable of antagonizing campylobacter jejuni and inhibiting expression of flaA gene of campylobacter jejuni
  • Lactobacillus plantarum capable of antagonizing campylobacter jejuni and inhibiting expression of flaA gene of campylobacter jejuni

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 screens lactic acid bacteria from Sichuan pickles

[0036] Samples were taken from Sichuan pickles and preserved in 30% glycerol. After homogenization, draw 1ml of kimchi suspension and add it to a test tube containing 9ml of sterilized physiological saline, oscillate evenly, and after gradient dilution, use the coating plate method to spread these diluted bacterial solutions on the surface added with the acid-base indicator olfactory cresyl violet On the MRS medium (Qingdao Haibo Biotechnology Co., Ltd. product) plate, carry out ordinary culture and anaerobic culture at a temperature of 37 ° C for 48 hours, observe the growth of the colony, and pick the bacteria that can make the olfactory cresyl purple in the medium turn yellow. The typical single colony, and carry out Gram staining on the single colony, pick the Gram-positive bacteria as the primary target strain, and then transfer to the MRS plate to separate and purify by streaking, until the pure lact...

Embodiment 2

[0037] Embodiment 2 uses the Oxford cup method to screen the lactic acid bacteria that can inhibit the growth of Campylobacter jejuni

[0038] Preparation of lactic acid bacteria fermentation supernatant: Inoculate the separated and purified lactic acid bacteria into MRS liquid medium at 2% by volume based on the volume of MRS liquid medium, culture at 37°C for 18 hours, and continuously culture for three generations in the same way, and then inoculate the lactic acid bacteria The bacterial suspension was centrifuged at 8000r / min and 4°C for 8min, the supernatant was absorbed, and sterilized by filtration with a 0.22μm water filter membrane to obtain the lactic acid bacteria fermentation supernatant.

[0039] Campylobacter jejuni (Campylobacter jejuni) NCTCl11168 strain (purchased from the American Type Culture Collection ATCC) on two media (i.e. Brooke's agar (product of Qingdao Haibo Biotechnology Co., Ltd.) and brain heart infusion broth medium ( Oxoid medium)) at a tempera...

Embodiment 3

[0044] Example 3 Tolerance experiment of Lactobacillus plantarum CGMCCNo.11445 to artificial intestinal juice and artificial gastric juice

[0045] Artificial gastric juice: Dissolve 3 g of pepsin (1:10000, Sigma, USA) in 1 L of normal saline (0.5%, w / v), adjust the pH to 2.0, pass the solution through a 0.45 / 0.22 μm sterile double-layer Pore ​​filter membrane, sterile storage for future use.

[0046] Artificial intestinal juice: Weigh 1g of trypsin (P-1500, Sigma, USA) and 0.3g of ox bile salt, dissolve in 1L of normal saline (0.5%, w / v), adjust the pH to 8.0, pass the solution through 0.45 / 0.22μm sterile double-layer microporous membrane, aseptically stored for future use.

[0047] PBS solution (phosphate buffered saline): Potassium dihydrogen phosphate 0.27g, potassium chloride 0.2g, disodium hydrogen phosphate dodecahydrate 2.85g, NaCl 8.5g, distilled water 1L, pH adjusted to 7.2, pass 0.45 / 0.22μm without Bacterial double-layer microporous membrane, sterilized at 121°C ...

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Abstract

The invention discloses lactobacillus plantarum. The Lactobacillus plantarum can antagonize campylobacter jejuni and inhibit expression of a flaA gene of the campylobacter jejuni, has the good acid resistance and bile salt resistance, can inhibit growth of the campylobacter jejuni in vitro and has the good adhesive capacity on the intestinal epithelial cells. A fermentation supernatant of the lactobacillus plantarum enables the expression amount of the flaA gene of the campylobacter jejuni to be reduced by 8.2 times compared with a control group in vitro, and synthesis of campylobacter jejuni flagella is effectively inhibited; the lactobacillus plantarum enables the expression amount of the flaA gene of the campylobacter jejuni to be reduced by 1.7 times compared with a control group in a cell model. The lactobacillus plantarum can be used for preparing medicine for preventing and treating diseases caused by the campylobacter jejuni and participate in preparing fermentation milk products and the like and has the wide application prospect.

Description

technical field [0001] The invention relates to a strain of lactobacillus plantarum which antagonizes Campylobacter jejuni and inhibits the expression of its flaA gene, and belongs to the technical field of microorganisms. Background technique [0002] Campylobacter jejuni is a Gram-negative bacterium that is widely distributed in nature and can be transmitted through animals, food, water, milk, etc. Infection in humans can result from ingestion of undercooked or contaminated chicken, beef, partially pasteurized milk, and contaminated water. In recent years, the infection rate of Campylobacter jejuni has been on the rise all over the world. In some developed countries, the number of cases of diarrhea caused by Campylobacter jejuni infection even exceeds that of Salmonella and Shigella, becoming the most common pathogen of diarrhea; in developing countries, Campylobacter jejuni is the most common pathogen of infectious diarrhea in infants and young children . When people a...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P31/04A23C9/123A23L1/29A23L1/218C12R1/25
CPCY02A50/30
Inventor 陈卫田丰伟张婷婷翟齐啸赵建新王刚张秋香刘小鸣张白曦张灏
Owner JIANGNAN UNIV
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