Antimicrobial Peptides
An antimicrobial peptide and bacterial infection technology, applied in the field of antimicrobial peptides, can solve problems such as easy inactivation and unstable structure of natural antimicrobial peptides, and achieve the effect of not easy inactivation, high antibacterial activity, and broad antibacterial spectrum
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Embodiment 1
[0019] Solid phase synthesis, cleavage, purification and identification of embodiment 1 antimicrobial peptide
[0020] 1.1 Solid-phase synthesis of antimicrobial peptides
[0021] The present invention utilizes the polypeptide synthesizer to adopt the solid-phase synthesis method to synthesize the antibacterial peptide:
[0022] 1) With DMF as solvent, the concentration of various α-amino acids protected by Fmoc is 0.25M, the concentration of HBTU solution and HOBt solution is 0.33M, the concentration of piperidine solution is 200ml / L, and the concentration of DIEA solution is 174.2ml / L.
[0023] 2) Weigh 0.05mmol of Fmoc-Ala-Wang resin (functional group content 0.33mmol / g) into a solid-phase reactor, add 8ml of DCM to swell overnight, and remove the solvent under reduced pressure. Add 8ml of piperidine solution with a concentration of 200ml / L, react at room temperature for 5min, and drain; then add 8ml of piperidine solution with a concentration of 200ml / L, react for 20min ...
Embodiment 2
[0036] Embodiment 2: the antibacterial activity assay of antimicrobial peptide
[0037] 1) Take 100 μl of the overnight cultured bacterial solution of different strains, dilute it in 10 ml of fresh medium, and cultivate it to the logarithmic growth phase (OD 570 0.6-0.8), and then diluted to OD value of 0.3 for use.
[0038] 2) Filter the prepared BSA solution with a 0.2 μm membrane, and the composition of the BSA solution is 20 mg of bovine serum albumin, 10 ml of water, and 1 μL of acetic acid.
[0039] 3) Dilute the polypeptide Phe-Lys-Ile-Gly-Gly-Phe-Ile-Lys-Lys-Leu-Trp-Arg-Ser-Leu-Leu-Ala (polypeptide 1 ) and Phe-Lys-Ile-Gly-Gly-Phe-Ile-Lys-Lys-Leu-Trp-Arg-Ser-Lys-Leu-Ala (polypeptide C). The final concentration is 10-320 μM.
[0040] 4) Dilute the bacterial solution with an OD value of 0.3 100 times for later use. Take a 96-well plate, add 180 μl fresh medium and 20 μl BSA solution to the blank control group; add 180 μl bacterial solution and 20 μl BSA solution to th...
Embodiment 3
[0050] Embodiment 3 hand sanitizer
[0051] Prepare the hand sanitizer containing the antimicrobial peptide polypeptide 1 of the present invention with the following composition:
[0052] components
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