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Application of bacillus subtilis in antagonism helicobacter pylori

A technology of Bacillus subtilis and Helicobacter pylori, which is applied in the directions of medical preparations, bacteria, and antibacterial drugs containing active ingredients to achieve strong antagonism, maintain intestinal microecological balance, and overcome the effects of large side effects.

Inactive Publication Date: 2011-07-27
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on the prevention and treatment of Helicobacter pylori infection by Bacillus subtilis at home and abroad

Method used

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  • Application of bacillus subtilis in antagonism helicobacter pylori
  • Application of bacillus subtilis in antagonism helicobacter pylori
  • Application of bacillus subtilis in antagonism helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. The 33 experimental strains were cultured in liquid at 37°C under anaerobic or aerobic conditions for 18-48 hours, centrifuged at 10,000 rpm for 30 min, and the supernatant was sterilized through a 0.22 μm filter.

[0026] 2. Pour the supernatant into a dialysis bag (molecular weight cut-off 3500 Da), clamp both ends with clips, and cover polyethylene glycol (PEG) around the outer surface of the dialysis bag. After about 4 hours, the supernatant is concentrated about 10 times , and collect the final concentrate for later use.

[0027] 3. Use a hole puncher to punch 4-6 holes on the brain heart infusion agar medium agar plate (the interval between holes is 20 mm), add 100 μL of culture supernatant and cell disruption solution to each hole, and place under microaerophilic conditions at 37°C After culturing for 48 hours, the diameter of the inhibition zone in each well was measured to analyze the inhibitory effect on the growth of Helicobacter pylori.

[0028] 4. T...

Embodiment 2

[0030] 1. Collect the culture supernatant of Bacillus subtilis CICC 23589 according to the method in Example 1 for subsequent use;

[0031] 2. Place the supernatant in a water bath at 45, 55, 65, 75, 85, 95, 105, and 115°C and heat for 5 min;

[0032] 3. Using the hole-diffusion method in Example 1, the effect of inhibiting Helicobacter pylori on the supernatant samples heated at the above-mentioned temperatures was determined.

[0033] 4. Experimental results such as figure 1 Shown: the abscissa indicates the heat treatment temperature (°C), and the ordinate indicates the antibacterial diameter circle (mm). From figure 1 It can be seen that the heat treatment below 95°C has basically no effect on the activity of the components in the supernatant that inhibit Helicobacter pylori. When the heat treatment temperature is higher than 95°C, the antibacterial components gradually lose their activity as the temperature rises.

Embodiment 3

[0035] 1. Collect the culture supernatant of Bacillus subtilis CICC 23589 according to the method in Example 1 for subsequent use;

[0036] 2. Prepare PBS (phosphate-buffered saline) buffer (20 mM), adjust pH to 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0; prepare Tris-HCl buffer (20 mM), adjust pH to 9.0, 10.0, spare;

[0037] 3. Put 2 mL of 10-fold concentrated supernatant of Bacillus subtilis CICC 23589 into a dialysis bag (molecular weight cut-off 3500 Da), and clamp both ends with dialysis bag clips;

[0038] 4. Dialyze the samples in each pH buffer solution overnight, change the buffer solution every 4 hours, end the dialysis at 48 hours, and collect the liquid in each dialysis bag;

[0039] 5. The antagonistic Helicobacter pylori activity of each collected sample was determined by punching diffusion method, and the diameter of the inhibition zone was measured;

[0040] 6. Experimental results such as figure 2 Shown: the abscissa indicates the pH value, and the ordinate ...

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Abstract

The invention relates to an application of bacillus subtilis in antagonism helicobacter pylori, wherein substances capable of restraining the growth of helicobacter pylori can be generated when the shake cultivation is performed on the bacillus subtilis for 9 hours at 37 DEG C and at a speed of 180 rpm (revolutions per minute) in a nutrient broth liquid culture medium, thereby efficiently restraining the growth of the helicobacter pylori; when the bacillus subtilis is cultivated for 18 hours, the antibiotic ability is maximum; and antagonistic proteins are acquired by purifying the substances, thereby preparing a preparation for treating gastrointestinal tract diseases. The invention has the following technical effects: the bacillus subtilis from probiotics has a function of maintaining the micro ecological balance of an intestinal tract; the bacillus subtilis culture has stronger antagonism to the helicobacter pylori; and compared with the antibiotic treatment method used for conventionally treating the helicobacter pylori, the bacillus subtilis can be used for overcoming the defects that the side effect by using antibiotic treatment method is strong and the symptoms are easily reappeared after the disease is treated, and the like.

Description

technical field [0001] The invention relates to an application of a bacillus subtilis, in particular to an application of a bacillus subtilis in antagonizing Helicobacter pylori. Background technique [0002] . In 1983, Marshall and Warren successfully isolated Helicobacter pylori for the first time. Many studies have confirmed that it is the main cause of chronic gastritis and duodenal ulcer, and is a potential factor leading to gastric cancer. [0003] Currently, the international standard triple therapy (standard triple therapy) is widely used to treat Helicobacter pylori infection. That is to say, when bismuth, metronidazole and tetracycline or ampicillin are used in combination, the eradication rate of Helicobacter pylori is 50-90%. This therapy has serious side effects, and may lead to the generation of drug-resistant strains of Helicobacter pylori at the same time, thereby further increasing the difficulty of treatment. In addition, under the induction of antibioti...

Claims

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Application Information

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IPC IPC(8): A61K35/74A61K38/00C12N1/20A61P1/04A61P31/04C12R1/125A61K35/742
Inventor 魏华曾明喻刚许恒毅程波财徐锋
Owner NANCHANG UNIV
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