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Lactic acid bacterium preparation and application thereof in treatment of gastric ulcer

A technology for lactic acid bacteria and gastric ulcer, applied in the direction of bacteria, microbe-based methods, and resistance to vector-borne diseases, etc., to achieve effective proliferation, improve curative effect, and strong tolerance

Active Publication Date: 2017-02-08
JINAN KANGDUOBAO BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(4) must actively compete for nutrients in the host

Method used

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  • Lactic acid bacterium preparation and application thereof in treatment of gastric ulcer
  • Lactic acid bacterium preparation and application thereof in treatment of gastric ulcer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 Lactobacillus fermentum KDB-8 isolation screening and identification:

[0025] 1. Screening samples

[0026] Naturally fermented sauerkraut juice was collected from the countryside of Sujiatun, Shenyang as an isolated sample. The sauerkraut juice has no pollution in appearance, pure taste, and has been fermented for more than one year. It is filtered and used for later use.

[0027] 2. Preliminary screening of lactic acid bacteria

[0028] Take 5 mL of filtered sauerkraut juice and add 45 mL of sterile water, mix well, spread on the MRS plate with 0.5% calcium carbonate, incubate at 37°C for 48 hours, isolate and culture the strains with transparent circles, and obtain 42 strains of lactic acid bacteria.

[0029] 3. Screening of antibacterial lactic acid bacteria

[0030] 1) Preparation of lactic acid bacteria liquid: inoculate 32 strains of lactic acid bacteria obtained from primary screening into 100mL MRS liquid medium, and culture at 37°C for 48 hours...

Embodiment 2

[0039] Example 2 Lactobacillus fermentum KDB-8 acid resistance, bile salt resistance experiment

[0040] 1. Gastric acid resistance test method:

[0041] Get 40ml of activated Lactobacillus fermentum KDB-8 bacterial suspension and add it to a 50ml centrifuge tube, centrifuge (5000g, 5min) to collect the thalline, wash twice with PBS buffer, add 40ml artificial gastric juice (take dilute hydrochloric acid) to the centrifuge tube Add 16.4ml of water and shake to dilute to 1000ml, adjust the pH to 2.0 with concentrated hydrochloric acid or 10% NaOH, sterilize at 121°C for 30min, add pepsin at a ratio of 1g / 100ml in a sterile room, incubate in a water bath at 37°C for 2h, Shake once every 30 minutes, take samples at 0h and 2h for counting viable bacteria, and use the 0h sample as a control to calculate the survival rate of the 2h sample bacteria, survival rate = (the number of viable bacteria at 2h / the number of viable bacteria at 0h) ×100%.

[0042] 2. Test method for bile salt...

Embodiment 3

[0048] Embodiment 3 Lactobacillus fermentum KDB-8 is to the removal effect of cholesterol

[0049] This experiment refers to the method of Brashears et al. (1998) with slight improvements.

[0050] The activated Lactobacillus fermentum KDB-8 strain was inoculated in the MRS medium and cultured overnight at 37°C as the seed solution.

[0051] Prepare MRS medium containing 5% (v / v) egg yolk liquid, take 2ml of medium, centrifuge at 5000r / min for 7min, take 1ml of supernatant and put it in a centrifuge tube, add 9ml of absolute ethanol, shake for 5min, 10000r Centrifuge for 10 min at 1 / min, take 2 ml of supernatant and add 2 ml of P-Fe-S reagent, mix and react in ice bath for 30 min, measure OD550, and calculate the initial content of cholesterol in the medium.

[0052] Then, inoculate the seed liquid of Lactobacillus fermentum KDB-8 into the above-mentioned MRS medium, and after culturing at 37°C for 24 hours, measure the final content of cholesterol in the medium again, and ca...

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Abstract

The invention relates to the technical field of microbiological screening and application and in particular provides a novel lactic acid bacterium preparation. The lactic acid bacterium preparation can be widely applied to treatment of gastric ulcer, the curative effect can be obviously improved, the cure rate of the ulcer is 90%, the eradication rate of helicobacter pylori is 84%, and the lactic acid bacterium preparation is high in tolerance to treatment and suitable for clinical application and popularization. The lactobacillus fermenti selected in the lactic acid bacterium preparation has obvious inhibitory effects on escherichia coli, salmonella and staphylococcus aureus, particularly has the strongest inhibitory effect on helicobacter pylori, and the diameter of an inhibition zone is 23mm. The lactobacillus fermenti has high tolerance to gastric acid and bile salt, and can realize effective multiplication in a gastric acid environment. The removal rate of lactobacillus fermenti KDB-8 on cholesterol is 93.2%; the survival rate of 98.5% can be maintained in 1.2mmol / L H2O2; and the scavenging rate of fermented supernatant fluid and cell-free extract on hydroxyl radicals is respectively 95.1% and 90.4%, and an undesirable technical effect is achieved.

Description

technical field [0001] The invention relates to the technical field of microorganism screening, and specifically provides a lactic acid bacteria preparation and its application in the treatment of gastric ulcer. Background technique [0002] The concept of probiotics comes from the Greek word "bacteria beneficial to life". Probiotics are usually the general term for active beneficial microorganisms (bacteria or yeast) that colonize in the intestinal tract and reproductive system of animals and can produce definite health effects. The FAO / WHO "Guidelines for the Evaluation of Food Probiotics" clearly stipulates that food probiotics refer to "living microorganisms that are beneficial to human health after taking a certain amount". The effectiveness of probiotics is closely related to the number of live bacteria and the performance of the strain itself. relevant. In my country, probiotics are also called microecological regulators, prebiotics, etc. Probiotic health food clearl...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61K9/19A61P1/04C12R1/225
CPCA61K35/747C12N1/20Y02A50/30
Inventor 尤升波郭学建刘凤彩张静孙玉亮张娟
Owner JINAN KANGDUOBAO BIOTECH CO LTD
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