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Application of ring(L-tyrosine-L-proline) in preparation of bacterial quorum sensing system inhibitors

A technology of quorum sensing system and tyrosine, which is applied in the application field of preparing bacterial quorum sensing system inhibitors, can solve problems such as drugs that have not yet been seen, and achieve the effect of no drug resistance and simple separation methods

Pending Publication Date: 2019-10-15
FUJIAN NORMAL UNIV
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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 bacterial QS, and no related drugs have been seen on the market.

Method used

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  • Application of ring(L-tyrosine-L-proline) in preparation of bacterial quorum sensing system inhibitors
  • Application of ring(L-tyrosine-L-proline) in preparation of bacterial quorum sensing system inhibitors
  • Application of ring(L-tyrosine-L-proline) in preparation of bacterial quorum sensing system inhibitors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Separation, purification and structural identification of ring (L-tyrosine-L-proline)

[0027] 1. Extraction of fermentation products

[0028] Penicillium chrysogenum DXY-1 (Penicillium chrysogenum DXY-1; its preservation number is: CCTCC NO: M2018731; preservation time: October 31, 2018; preservation address: China Type Culture Collection Center, the strain has been Disclosed in the application with the patent number 201910398145.4) after activation of the plate, scrape an appropriate amount of mycelium and inoculate it in the culture solution, and culture it with shaking at 28°C and 150r / min for 72 hours to obtain the seed culture solution. Then the seed liquid was transferred to a 1000 mL Erlenmeyer flask filled with 400 mL of Fungus No. 5 culture medium with a 5% inoculation amount, and cultured with shaking under the same conditions for 7 days to obtain a fermentation liquid. Use a homogenizer to crush the mycelium balls in the fermentation broth, then a...

Embodiment 2

[0036] Example 2 Cyclic (L-tyrosine-L-proline) inhibition of Pseudomonas aeruginosa PA01 virulence factor synthesis analysis

[0037] 1. Detection of pyocyanin production

[0038] Activate Pseudomonas aeruginosa PA01 to log phase and dilute to OD 600 ≈0.05, divided into 4 groups, 3 groups were parallel experiments, each group was added with different concentrations of compounds, the blank control was methanol, and the positive control was 0.2 mg / mL azithromycin, cultured with shaking at 37°C for 12 hours. Centrifuge to remove bacteria, take 5mL of supernatant and add 3mL of chloroform for extraction, transfer the chloroform phase into a new centrifuge tube and add 1mL of 0.2mol / L hydrochloric acid, mix well and extract, collect the upper aqueous phase by centrifugation, and measure the wavelength with a microplate reader OD value at 520nm, see the results figure 1 .

[0039] 2. Detection of protease production

[0040] To activate Pseudomonas aeruginosa PA01 overnight, ino...

Embodiment 3

[0046] Embodiment 3 ring (L-tyrosine-L-proline) inhibits the analysis of Pseudomonas aeruginosa biofilm formation

[0047] SEM imaging of biofilm: activate Pseudomonas aeruginosa PA01 overnight, inoculate 1% PA01 in liquid LB medium, put sterile glass slides in six-well plate and add 3mL bacterial solution, add ring (L-casein Amino acid-L-proline), positive control 0.2mg / mL azithromycin and blank control methanol, cultured at 37°C for 48h. Aspirate the bacteria solution, wash the slides with sterile PBS, fix with 2.5% glutaraldehyde at room temperature for 24h, and then wash with PBS three times to remove excess glutaraldehyde. Dehydrate with gradient concentrations (50%, 70%, 80%, 90%) of ethanol for 15 minutes, and finally dehydrate with 100% ethanol three times, each time for 10 minutes. Spray gold observation, the results see figure 2 .

[0048] figure 2 Middle CK is the PA01 biofilm treated with blank control methanol, the surface is dense, and the cells are complet...

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Abstract

The invention discloses application of a ring(L-tyrosine-L-proline) in preparation of bacterial quorum sensing system inhibitors. The invention aims to provide a novel bacterial QS inhibitor ring(L-tyrosine-L-proline), the content of the ring(L-tyrosine-L-proline) is relatively high in the metabolites marine fungi, and the ring(L-tyrosine-L-proline) can be extracted on a large scale, or can be obtained by artificial synthesis. Therefore, according to the present invention, the ring(L-tyrosine-L-proline) has the advantages of simple separation method and no drug resistance, and is suitable forlarge-scale preparation.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to the application of ring (L-tyrosine-L-proline) in the preparation of bacterial quorum sensing system inhibitors. Background technique [0002] The World Health Organization (WHO) pointed out in a report published in 2014 that the onset and infection of "super bacteria" with strong drug resistance is spreading and becoming more serious year by year, and the survival of human beings is greatly threatened. Antimicrobial resistance of bacteria has become a major problem of global concern. Therefore, finding new targets is a new strategy for the development of new antibacterial drugs in the field of biomedicine. Quorum sensing (QS) is a communication system between bacterial cells, which can regulate the survival and pathogenicity of bacteria, which has attracted close attention from pharmacists and has become an important target for the development of new antibacterial drugs . T...

Claims

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Application Information

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IPC IPC(8): A61K38/05A61K31/4985A61P31/04C12P21/04C07K5/12C12R1/82
CPCA61K38/05A61K31/4985A61P31/04C12P21/02C07K5/12
Inventor 朱虎王颖璐常爱平李丽鱼晓丹何侨妹戴晓芸
Owner FUJIAN NORMAL UNIV
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