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Application of benzyl benzoate in preparation of bacterial colony induction activity inhibitors

An activity inhibitor, the technology of benzyl benzoate, which is applied in the application field of benzyl benzoate in the preparation of bacterial quorum sensing activity inhibitors, can solve problems such as drugs that have not yet been seen, and achieve controllable fermentation and culture conditions and a high preparation process. The effect of simple process and stable yield of metabolites

Active Publication Date: 2018-01-26
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report that this compound has the activity of inhibiting the quorum sensing activity of Violet Bacillus and Pseudomonas aeruginosa, so there is no related drug on the market

Method used

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  • Application of benzyl benzoate in preparation of bacterial colony induction activity inhibitors
  • Application of benzyl benzoate in preparation of bacterial colony induction activity inhibitors
  • Application of benzyl benzoate in preparation of bacterial colony induction activity inhibitors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Purification and structural identification of embodiment 1 benzyl benzoate

[0029] (1) Preparation of crude extract: inoculate Pacific bacillus XC22919 (Oceanobacillus sp.XC22919) into slant medium (ie: LB seawater solid medium: peptone 10g / L, yeast extract 5g / L, NaCl 10g / L, sea salt 33.3g / L, agar 18g / L, the solvent is deionized water, pH 7.4; 121°C, sterilized for 20min), cultivated overnight in a constant temperature incubator at 30°C to obtain slant bacteria; then inoculate the slant bacteria to the seeds Culture medium (that is, LB seawater liquid medium, remove the agar in LB seawater solid medium, and the others are the same), in a constant temperature shaker at 30°C, vibrate at 180rpm overnight to obtain seed liquid; The inoculum amount was inoculated into LB seawater liquid medium (peptone 10g / L, yeast extract 5g / L, NaCl 10g / L, sea salt 33.3g / L, solvent was deionized water, pH 7.4; 121°C, sterilized for 20min ), shaking and culturing for 3 days at 180 rpm in a c...

Embodiment 2

[0036] The effect of embodiment 2 benzyl benzoate on inhibiting the growth of Chromobacter violaceum and the production of purple pigment

[0037] (1) Experimental method: Inoculate Chromobacterium violaceum CV026 containing exogenous signal molecule (Chromobacterium violaceum, donated by Professor David C. Rowley, University of Rhode Island, USA) into LB liquid medium, culture overnight at 30°C, and obtain Violet bacteria CV026 liquid.

[0038] After diluting the violaceum bacteria CV026 liquid with fresh LB liquid medium at a volume ratio of 1:100, 10 mL per bottle. Add different final concentrations (0, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / mL, 60 μg / mL, 70 μg / mL, 80 μg / mL) benzyl Added in the form of methanol solution), and methanol was used as a negative control. Place it on a shaker at 30°C and 150rpm for about 18h, take 1mL of the bacterial solution, centrifuge for the first time at 12000rpm for 10min, discard the supernatant, add 1mL of DMSO, vortex and shake...

Embodiment 3

[0040] Effect of embodiment 3 benzyl benzoate on the growth of Pseudomonas aeruginosa PAO1 and the output of pyocyanin

[0041] (1) Experimental method: Pseudomonas aeruginosa (Pseudomonas aeruginosa) PAO1 was inoculated into LB medium, cultured overnight at 37° C. to obtain Pseudomonas aeruginosa PAO1 bacterial liquid.

[0042] Pseudomonas aeruginosa PAO1 bacteria liquid was used fresh PB liquid culture medium (peptone 20g / L, magnesium chloride 1.4g / L, potassium sulfate 10g / L, solvent was deionized water, pH natural; 121 ℃, sterilization 20min) with After dilution by volume ratio of 1:100, each bottle is 10mL. Benzyl benzoate (in the form of 50 mg / mL methanol solution) was added at different final concentrations (0, 20 μg / mL, 40 μg / mL, 60 μg / mL, 80 μg / mL, 100 μg / mL, 120 μg / mL) to Methanol served as a negative control. Place it on a shaker at 37°C and 150rpm for about 24h. Take 6mL of bacterial liquid, add 3mL of chloroform for extraction; after extraction, the chloroform l...

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Abstract

The invention discloses an application of benzyl benzoate in preparation of bacterial colony induction activity inhibitors; in the method, an oceanobacillus sp. XC22919 fermented product is used for separation and extraction, and in a concentration range of 0-60 [mu]g / mL, bacterial growth of chromobacterium violaceum CV026 is not inhibited, but generation of chromobacterium violaceum purpurin canbe significantly decreased; and with gradual increase of the concentration, the effect of inhibiting generation of the purpurin is stronger; the effect of the compound in inhibiting colony induction is in a concentration dependence. In a concentration range of 0-100 [mu]g / mL, bacterial growth of pseudomonas aeruginosa PAO1 is not affected, but the expression of virulence factors of pseudomonas aeruginosa can be significantly reduced, the expression of pyocyanin, elastase, proteinase, biological body membranes and other virulence factors can be significantly reduced, and the effect of the compound in inhibiting colony induction is in the concentration dependence.

Description

(1) Technical field [0001] The invention relates to a compound with bacterial quorum sensing inhibitory activity isolated and purified from marine-derived Oceanobacillus sp. XC22919 (Oceanobacillus sp. XC22919) through fermentation, identification of its structure and application thereof. (2) Background technology [0002] It is traditionally believed that there is information exchange only between eukaryotic cells, but not between lower microorganisms. However, it was later discovered that during the process of growth and reproduction, bacteria also secrete some specific signal molecules into the surrounding environment according to changes in the environment, and detect changes in themselves or other microbial populations in the environment by sensing changes in these signal molecules Scientists call this signaling molecule an autoinducer (AI). Bacteria continuously secrete signal molecules to the outside world. When a threshold is reached in a certain period of time, bac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62A61K31/235A61P31/04C12R1/01C12R1/385
Inventor 王鸿严银春陈小春陈建伟章华伟
Owner ZHEJIANG UNIV OF TECH
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