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Bacillus subtillus degrading bacterial colony sensing signal and use of bacillus subtillus degrading bacterial colony as antiseptic

A technology of Bacillus subtilis and quorum sensing signal, which is applied in the field of hygiene, can solve the problems restricting the antibacterial application of Bacillus subtilis, and achieve the effects of being suitable for popularization and use, avoiding drug resistance problems, and using safely

Inactive Publication Date: 2010-12-29
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This restricts the antibacterial application of Bacillus subtilis to a certain extent

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Screening, isolation and purification of Bacillus subtilis:

[0026] The 2216E plate was cultured at 30°C for 2-3 days, separated and purified by streaking several times, inoculated on the slant, and stored at 4°C for future use.

Embodiment 2

[0028] Preliminary screening experiment of Bacillus subtilis degrading bacterial signal molecules:

[0029] In the preliminary screening experiment, the signal molecule was OOHL molecule, and the indicator bacteria was WCF47.

[0030] 96-well plate primary screening:

[0031] 1) Pick the bacteria to be tested from the plate and culture them in 96-well culture plate A with 200 μl of the corresponding liquid medium until late logarithmic growth, and set up a negative control (LB+AHL) and a positive control (Bt+AHL).

[0032] 2) Add 100 μl MM liquid medium and 0.2 μl signal molecule AHL (1 mg / ml) to each well of the 96-well culture plate B, and use a pipette gun to take 100 μl of the bacteria to be tested from the 96-well culture plate A into the 96-well culture into the corresponding wells of Plate B, and mix well. Co-cultivate at 30°C for 4hr.

[0033] 3) Add 130 μl MM liquid medium, 20 μl indicator bacteria WCF47 bacterial solution, and 1 μl X-Gal to each well of the 96-wel...

Embodiment 3

[0037] Re-screening experiment of Bacillus subtilis degrading bacterial signal molecules:

[0038] Re-screening of agar strips:

[0039] Cut the MM medium plate coated with X-gal into strips, add the mixture of test bacteria and AHLs extract to one strip, add indicator bacteria WCF47 to the other strip, and observe the experimental results after culturing at 30°C for 18 hours . LB or ddH 2 The mixture of O and AHL was used as a negative control, and the mixture of AiiA active bacteria B. thuringiensis culture and AHL was used as a positive control.

[0040] Experimental results: According to the color reaction, the negative control is blue, and the positive control and Bacillus subtilis are white, indicating that Bacillus subtilis has the activity of degrading AHLs signal molecules.

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PUM

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Abstract

The invention belongs to the field of hygiene and relates to a bacillus subtillus degrading bacterial colony sensing signal and use of bacillus subtillus degrading bacterial colony as an antiseptic, in particular to bacillus subtillus degrading gram-negative bacterial colony sensing signal and the formation of a bacterial biofilm of the bacillus subtillus degrading gram-negative bacterial colony. In the use of the degrading bacterial colony sensing signal, a proper concentration of the bacillus subtillus is 5*102 to 5*104cfu / ml. The bacillus subtillus is further used as antiseptic to inhibit the bacteria such as pseudomonas, agrobacterium, enterobacteriaceae and vibrionaceae. The antiseptic can be ointment for external use, liquid preparation for external use, oral tablet, oral liquid, disinfectant, clearing agents or water quality improving agent. The consistency of the bacillus subtillus serving as the antiseptic is 105 to 106 cfu / g. The use of the bacillus subtillus provided by the invention and the antiseptic inhibit the formation of biofilm of disease-causing bacteria and the growth of bacteria by using microbial quorum-sensing inhibition instead of preventing and sterilizing bacteria by compound which is adopted by most of the conventional antiseptic; therefore, the problem of drug resistance in the antimicrobial process is avoided.

Description

technical field [0001] The invention belongs to the field of hygiene, and relates to a Bacillus subtilis degrading bacterial quorum sensing signal and its application as an antibacterial agent. Background technique [0002] Bacillus stibillus is a microorganism isolated and purified from the culture environment. The physiological and biochemical characteristics of the strain were identified according to Bergey's manual. The Bacillus subtilis was nearly round on the ordinary medium, opaque, dry on the surface, wrinkled around, and the edges were uneven. Gram staining was positive under the microscope, and the bacteria were rod-shaped. . [0003] Also has the following biochemical properties: Mannitol produces acid +, arabinose produces acid +, glucose produces gas -, forms indole -, V.P test +, phenylalanine deamination -, V.p culture final pH 5.8, glucose produces acid +, starch hydrolysis+, citrate utilization+, catalase+, nitrate reduction+, NaCl, KCl need+, 10% NaCl gro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74A01N63/00C02F1/50C11D3/48C11D9/50A01P1/00A01P3/00A61P31/02A61P31/04A61K35/742
Inventor 丁贤殷波周世宁江世贵
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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