Antibacterial peptide and application thereof in preparation of medicines or cosmetics
An antifungal drug and antibacterial peptide technology, applied in cosmetics, cosmetic preparations, antifungal agents, etc., can solve the problems of no research and no screening of antibacterial peptides, and achieve the effect of significant inhibitory activity and low toxicity
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Embodiment 1
[0022] Example 1 Screening of anti-Trichophyton rubrum bactericidal peptides
[0023] 1. Bacteria culture
[0024] The standard strain of Trichophyton rubrum ATCC®28188 was planted on Sabouraud dextrose agar SDA plates, cultured in the dark at 27°C, picked a single colony and put it into a shaker flask of 250mL PDA medium, 150r / min, cultured in the dark at 27°C, Then, centrifuge at 3000 r / min, 4°C for 10 min, and collect the bacterial cells to harvest the bacterial cells for later use.
[0025] 2. Lipid extraction
[0026] 8 g of the collected bacterial cells were washed repeatedly with PBS, centrifuged, resuspended in 60 mL Tris-HCl buffer (25 mmol / L, pH7.5), ultrasonically disrupted, and after centrifugation, 180 mL of chloroform-methanol (1:2 , v / v) mixed solvents, mixed and stirred for 100 min. After that, add 40 mL of chloroform and 40 mL of distilled water, and continue stirring for 30 min. The chloroform phase was separated with a separatory funnel, and the solvent ...
Embodiment 2
[0036] The mensuration of embodiment 2 antimicrobial peptide minimum inhibitory concentration
[0037] The minimum inhibitory concentrations (Minimalinhibitoryconcentrations, MICs) of antimicrobial peptides against indicator bacteria were determined by microdilution method.
[0038] The strains tested were: (1) Escherichia coli ATCC25922; (2) Trichophyton mentagrophytes, NRR00060 of Beijing Huayueyang Biotech Co., Ltd.; (3) Trichophyton rubrum ATCC®28188. The strains were cultured according to conventional culture methods. After culturing to the logarithmic phase, the bacterial cells in the logarithmic phase were collected by centrifugation (3000 r / min, 4°C, 10 min), washed with sterilized PBS, and resuspended until the OD600 was about 0.3. The peptide was prepared into a stock solution of 1024 µg / mL with sterilized PBS, sterilized by filtration with a 0.22 µm filter membrane, and serially diluted with sterilized PBS. Add 100 µL medium, 100 µL test bacteria solution and 50 µ...
experiment example 3
[0042] Experimental example 3 The effect of antimicrobial peptides in the treatment and prevention of skin ringworm
[0043] Trichophyton rubrum was activated and inoculated in SDA medium, cultured at 30°C for 7 days, eluted with sterile normal saline, shaken and mixed well, and adjusted the concentration of the mixed bacterial suspension to 1.0×10 8 CFU / mL. Anesthesia was injected intraperitoneally, and the back and sides of guinea pigs were sheared and depilated respectively to form a hairless area of about 2.5×2.5 cm each. After the hairless area was polished with sterile sandpaper until spot-like bleeding, 100 μL of bacterial suspension was evenly applied. On the 3rd day of inoculating the bacterial solution, 25 guinea pigs successfully modeled were numbered respectively and divided into 5 groups according to the random number table method (negative control group / 5% ethanol solvent group / 8 μg / mL antimicrobial peptide / 32 μg / mL antimicrobial peptide / positive control gro...
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