Application of hydroxyphenethyl alcohol for preparing medicine for restraining bacterium quorum sensing
A technology of p-hydroxyphenethyl alcohol and quorum sensing, applied in the directions of antibacterial drugs, active ingredients of hydroxyl compounds, etc., can solve problems such as the application of hydroxyphenethyl alcohol that has not been reported yet, and achieve the effect of inhibiting bacterial quorum sensing.
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Embodiment 1
[0044] Example 1 P-Hydroxyphenyl Alcohol Inhibits Activity Test of Violet Bacillus Quorum Sensing
[0045] 1. Experimental principle
[0046] (1) Bacterial quorum sensing (QS) indicator bacteria Violaceum CV026 is the signal molecule C 6 The cviI deletion mutant of HSL synthase gene, exogenously added C6HSL signal molecule, can induce its quorum sensing to produce violacein. If a compound has bacterial quorum sensing inhibitory activity, the compound will reduce violacein production.
[0047](2) The synthesis of violacein is strictly regulated by the bacterial quorum sensing system of Violaceum. By analyzing the growth of Violaceum and the production of violacein, it can be judged whether a certain compound has bacterial quorum sensing inhibitory activity.
[0048] 2. Experimental method
[0049] 2.1 Violet bacteria plate screening method
[0050] Violaceum CV026 was activated overnight. Cool 20 mL of melted solid LB medium to 28 °C, add 20 μL of kanamycin, 100 μL of si...
Embodiment 2
[0058] Example 2 P-hydroxyphenylethanol inhibits the activity test of Pseudomonas aeruginosa quorum sensing
[0059] 1. Experimental method
[0060] 1.1 Pyocyanin production
[0061] Activate P. aeruginosa PA01 to logarithmic phase and dilute to OD 600 0.05, and then divided into 7 groups on average (blank control group, positive control group and 5 p-hydroxyphenylethanol groups with different concentrations); wherein, 5 groups were added p-hydroxyphenylethanol respectively to form different concentration gradients of 0.1mg / mL , 0.2mg / mL, 0.3mg / mL, 0.4mg / mL and 0.5mg / mL p-hydroxyphenylethanol solutions; in addition, methanol was used as a blank control group, and 0.2mg / mL azithromycin was used as a positive control group; 37°C , shaking culture for 12h. Centrifuge to remove Pseudomonas aeruginosa cells, take 5mL supernatant and add 3mL chloroform for extraction, transfer the chloroform phase into a new eppendorf tube, add 1mL of 0.2mol / L hydrochloric acid, mix well and ex...
Embodiment 3
[0068] Embodiment 3 p-Hydroxyphenyl alcohol inhibits the analysis of the biofilm formation of Pseudomonas aeruginosa
[0069] 1. Experimental principle
[0070] The quorum sensing system of bacteria plays an important role in the formation and structural stability of biofilms. Inhibiting the quorum sensing system of P. aeruginosa PA01 can effectively reduce the formation of biofilms and improve drug sensitivity.
[0071] 2. Experimental method
[0072] 2.1 SEM imaging of biofilm
[0073] P. aeruginosa PA01 was activated overnight. The activated Pseudomonas aeruginosa P.aeruginosaPA01 was inoculated into the liquid LB medium, and the inoculation amount of the Pseudomonas aeruginosa P.aeruginosa PA01 was 1vol% of the volume of the liquid LB medium; Put sterile glass slides, then add p-hydroxyphenylethanol (p-hydroxyphenylethanol group), 0.2 mg / mL azithromycin (positive control group) and methanol (blank control group) respectively, after static incubation at 37°C for 48 hou...
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