A kind of termite antimicrobial peptide and its application
A technology of antimicrobial peptides and termites, applied in the field of bioengineering, can solve the problems that antimicrobial peptides have not yet been discovered, and achieve the effect of normal cell toxicity, small molecular weight, and strong bactericidal effect
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Embodiment 1
[0024] The extraction of embodiment 1 antimicrobial peptide
[0025] 1. Preparation of termite samples
[0026] After the termite sample is dried, take 5-10 g of the sample, pulverize it with an electric pulverizer, and set it aside.
[0027] 2. Degreasing treatment
[0028] Wrap the prepared termite powder with a filter paper tube, put it into the extraction tube of the Soxhlet extractor, connect it to a fat flask that has been dried to constant weight, and add ether or petroleum ether into the bottle from the upper end of the condenser tube of the extractor Fill the 2 / 3 of the volume with condensed water, immerse the bottom bottle in hot water at 65°C, heat it in a water bath, gently insert a small ball of absorbent cotton into the upper opening of the condenser tube, extract for 6-12 hours, and then put it in the filter paper tube The samples were fully dried at 50°C.
[0029] 3. Ethanol extraction
[0030] According to the ratio of solid-liquid ratio of 1:30, add 60% e...
Embodiment 2
[0042] Bacteriostatic effect detection
[0043] The bacteriostasis was detected by the Oxford cup method. The bacteria to be tested were Escherichia coli (ATCC 25922), Staphylococcus aureus (ATCC 29213), Klebsiella pneumoniae strain WL1309, Salmonella enteritidis (ATCC 13076 ), Bacillus subtilis (ATCC 11060) for antibacterial experiments, use a sterile pipette to draw 0.1mL of the prepared bacterial suspension, add them to the solid plate of the culture medium corresponding to the tested bacteria, and use a sterile coating Spread the cloth stick evenly to make a flat plate with bacteria (or mix the bacteria with a medium with a suitable temperature to make a pouring flat plate). Under aseptic conditions, put the sterilized Oxford cup into the flat plate with tweezers, and extract The antimicrobial peptide material that obtains, detects whether it has antibacterial zone (transparent) to occur, and its antibacterial effect sees figure 1 ,in, figure 1 a is the bacteriostatic di...
Embodiment 3
[0046] Minimal Inhibitory Concentration Test
[0047]The purified termite antimicrobial peptide extracted in Example 1 was diluted with different decreasing concentrations to obtain concentrations of 100mg / L, 90mg / L, 80mg / L, 70mg / L, 60mg / L, 50mg / L, 40mg / L, 30mg / L, 20mg / L and 10mg / L antimicrobial peptide dilutions were respectively used to detect the minimum inhibitory concentration of Escherichia coli and Klebsiella pneumoniae, among which the antibacterial peptide dilution The minimum inhibitory mass concentration is the diameter of the circle above 10mm; the experiment is repeated 3 times, and the average value is calculated, and the test results are shown in figure 2 ;in, figure 2 a is the bacteriostatic diagram of Escherichia coli; figure 2 b is the bacteriostatic diagram of Klebsiella pneumoniae, through figure 2 According to calculations, the minimum inhibitory concentration of the antibacterial peptide to Escherichia coli is 16.3 mg / L, and the minimum inhibitory ...
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Abstract
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