Bacterial quorum sensing inhibitor and antibacterial application thereof
A quorum sensing system and bacterial technology, applied in the field of microorganisms, can solve the problems of no antibacterial agents and low antibiotics
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Embodiment 1
[0019] Example 1: Purification and identification of the traditional Chinese medicine notopterygium falcarindiol
[0020] Preparation method of methanol crude extract: grind notopterygium rhizomes and weigh 10 g of powder, extract each 2.5 g of powder with 200 mL of methanol, sonicate for 1 h and soak for 24 h; filter paper to obtain the leachate, and evaporate the filtrate to dryness with a rotary evaporator and weighed; re-dissolved with 30 mL of methanol and then filtered to remove insoluble matter with a homemade cotton filter; evaporated the filtrate again and weighed.
[0021] The preparation method of the compound: after the above-mentioned extract is stirred evenly with an appropriate amount of silica gel H, it is separated by vacuum liquid chromatography column chromatography, and the eluent petroleum ether, dichloromethane, dichloromethane:methanol (100: 1), dichloromethane:methanol (50:1), dichloromethane:methanol (20:1), and methanol were washed separately, and eac...
Embodiment 2
[0028] Example 2: Quorum Sensing Inhibition Activity Test of Fakalindiol against Violet Bacillus
[0029]Violet bacteria screening model CV026 composition and method: Cool 19 mL of melted solid LB medium to 40°C, add 600 μL overnight cultured violaceum CV026 bacterial liquid, 15 μL signal molecule hexanoyl homoserine at a concentration of 500 μmol / L ester (C6-HSL) and 15 μL of kanamycin at a concentration of 30 mg / mL, and mix them into a Petri dish. After the plate is solidified, punch holes with a sterilized hole puncher, add 5 μL of compound to each hole, incubate in a 30°C incubator for 12 hours, and observe the growth of bacteria around the sample hole. If the compound is a quorum sensing inhibitor, it can inhibit the production of violacein, and it will appear as a turbid but opaque circle on the agar plate; the larger the turbid circle, the higher the activity of the compound.
[0030] Experimental results: compared with the positive control furanone C 30 (6.5 μg), 6.5...
Embodiment 3
[0031] Embodiment 3: Quorum sensing inhibitory activity test of anti-Pseudomonas aeruginosa of fakalindiol
[0032] Inhibition of Pseudomonas aeruginosa AHL quorum sensing system:
[0033] Components and methods of Pseudomonas aeruginosa detection model QSIS2: 17.5 mL of melted solid LB medium was cooled to 40°C, 2 mL of 60% sucrose was added, 250 μL of Pseudomonas aeruginosa QSIS2 cultured overnight, 8 μL signal molecule 3-oxolauroyl homoserine lactone (3-oxo-C12-HSL) at a concentration of 100 μmol / L, 200 μL of a viable bacterial stain red tetrazolium at a concentration of 5% (w / v) TTC and 32 μL of gentamicin at a concentration of 50 mg / mL, mix well and pour into a petri dish. After the plate is solidified, punch holes with a sterilized hole puncher, add 5 μL of compounds of different concentrations to each hole, place them in an incubator at 37°C, and observe the growth of bacteria around the sample holes. If the compound has quorum sensing inhibitory activity, a red bacte...
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