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A strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance

A technology of Lactobacillus plantarum and Lactobacillus, applied in bacteria, microorganisms, microorganisms and other directions, can solve the problems of complex causes of intestinal diseases, no effective measures for long-term sub-health state, and insufficient symptoms, so as to improve intestinal function. , the preparation method is simple, the effect of high bacterial survival rate

Active Publication Date: 2021-03-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the causes of intestinal diseases caused by the use of antibiotics are relatively complicated, the symptoms are not obvious enough, and there are no effective measures for the long-term sub-health state caused by it

Method used

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  • A strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance
  • A strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance
  • A strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance

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Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1: the determination of the growth generation time of different plant lactobacillus

[0035] The generation time refers to the time required for each cell division, and the specific growth rate is the largest in the logarithmic phase, so in the present invention, the bacterial generation time in the logarithmic growth phase of different Lactobacillus plantarum is measured under aerobic culture at 37°C.

[0036] The bacteria were inoculated in MRS medium with 2% inoculation amount, cultured at 37°C, and samples were taken every 0.5 h to measure the absorbance at 600 nm (OD600) of the bacteria solution. According to the law of bacterial growth, in the logarithmic growth period: A t =A 0 *2(t-0) / G, where A 0 Represents the initial number of bacteria when entering the logarithmic growth phase; A t Indicates the number of bacteria at time t; G represents the generation time of bacteria. Derivable formula: log 2 A t = log 2 A 0 +(1 / G)*t. It can be seen tha...

Embodiment 2

[0038] Embodiment 2: Determination of the survival rate of Lactobacillus plantarum in simulated gastrointestinal fluid in vitro

[0039] Prepare simulated gastrointestinal fluid, the simulated gastric juice is 6.2g / LNaCl, 2.2g / LKCl, 0.22g / L CaCl 2 , 1.2g / LNaHCO 3 , 0.3% pepsin, adjusted to pH 2.0 with 6M HCl; synthetic intestinal fluid 6.4g / LNaHCO 3 , 0.239g / L KCl, 1.28g / L NaCl and 0.1% pancreatin, adjusted to pH 7.4 with 6M HCl. After fully dissolving, filter and sterilize with a 0.22μm microporous membrane, dispense 5mL into each test tube, and refrigerate at 4°C for later use. Centrifuge the lactic acid bacteria cultured for 18 hours, resuspend and metabolize them with 0.9% normal saline for 1 hour, then insert them into the simulated gastric juice and mix them evenly, place them in an oscillating water bath at 37°C with a speed of 75r / min to simulate human gastrointestinal peristaltic digestion, and take them out after 2 hours . After centrifugation, simulated intestin...

Embodiment 3

[0045] Example 3: Determination of Adhesion Performance of Different Lactobacillus plantarum to Human Colon Cancer Cell HT-29

[0046] HT-29 cells were cultured in RPMI 1640 (10% fetal bovine serum, 1% antibiotics) medium, and the medium was changed every two days until the cells reached 80-90%. Digest the cells with 0.25% trypsin solution and adjust the cell concentration to 1 x 10 5 cells / mL, and inoculated in a six-well culture plate, put a 18×18mm sterile coverslip in the culture plate in advance, and place it at 37°C, 95% air / 5% CO 2 Culture in an incubator until a dense monolayer of cells grows. After washing the cells twice with PBS buffer (pH 7.2), add 1 mL of antibiotic-free RPMI medium and 1 mL of cultured and resuspended bacteria suspension in PBS to each well (adjust the total number of bacteria to 10 8 ), mixed evenly and incubated in a 5% carbon dioxide incubator, each strain had three parallels. After culturing for 2 hours, the culture plate was taken out, an...

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Abstract

The invention discloses a strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance, which belongs to the technical field of food. The present invention is to expand the cultivation of Lactobacillus plantarum (Lactobacillus plantarum) CGMCC12436, supplemented with skim milk, trehalose and sucrose, and prepare a biological preparation after freeze-drying. The live bacteria content of Lactobacillus plantarum CGMCC12436 in this preparation is greater than 10 9 CFU / g. The Lactobacillus plantarum CGMCC12436 provided by the present invention has a relatively low growth generation time, a high survival rate in simulated gastric juice and small intestinal juice in vitro, good adhesion to human HT-29 cells, and relatively low drug resistance in the drug sensitivity test. lower. The biological preparation made by the Lactobacillus plantarum can significantly restore the intestinal flora imbalance caused by ampicillin in mice, improve intestinal function, and has a good market application prospect.

Description

technical field [0001] The invention relates to a strain of Lactobacillus plantarum capable of regulating ampicillin-induced intestinal flora disturbance, belonging to the technical field of food. Background technique [0002] Gut flora is an important part of the human gut and plays a major role in intestinal metabolism and physiological functions. The balance of intestinal flora is affected by the host's physiology, food, drugs, and the interaction between intestinal flora. Various factors may cause intestinal flora disorder, resulting in an increase in conditional pathogenic bacteria, a decrease in the proportion of beneficial bacteria, and metabolic Reduced function, thus causing intestinal diseases such as antibiotic-associated diarrhea, irritable bowel syndrome and inflammatory bowel disease. The use of antibiotics will lead to intestinal symptoms such as a decrease in intestinal flora, a relative increase in drug-resistant bacteria, and metabolic disorders, which has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23C9/127A23L11/65A23L11/50C12R1/25
CPCA23C9/127A23C11/106C12N1/20C12R2001/25C12N1/205
Inventor 陈卫翟齐啸史瑛田丰伟赵建新张灏
Owner JIANGNAN UNIV
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