An Antimicrobial Peptide Derived from Amphioxus
A technology of antibacterial peptides and amino acids, applied in the direction of antibacterial drugs, antifungal agents, peptides, etc., can solve the problems of changing the permeability of cell membranes such as antibacterial peptides
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Embodiment 1
[0018] Embodiment 1: Screening and detection of antimicrobial peptides
[0019] Because the antimicrobial peptide gene generally has a long precursor peptide before it is processed into a mature peptide through post-translational processing, the sequence of the mature peptide part of the antimicrobial peptide is very different among species with similar evolutionary relationships, while the sequence of the precursor peptide The sequence is relatively conserved. Therefore, according to the characteristic that the signal peptide sequence of the precursor peptide encoded by the antimicrobial peptide gene between the species with close evolutionary relationship is highly conserved, select Styela clava, Ciona intestinalis, Myxineglutinosa, The antimicrobial peptide precursor peptide of horseshoe crab was used as a search condition to screen peptides from the amphioxus database.
[0020] The polypeptide sequences with high homology to the signal peptides of known antimicrobial pept...
Embodiment 2
[0031] Example 2: Destruction effect of AP22 polypeptide on bacterial cell wall
[0032] Add antibacterial peptide AP22 polypeptide at a concentration of 5 μg / ml to the culture medium of Escherichia coli O1 strain, incubate at 37°C for 60 minutes, collect the bacterial liquid, centrifuge at 4000r / min for 10 minutes, discard the supernatant, and replace with 0.1mol / L phosphoric acid Saline buffer (pH 7.0) for washing.
[0033] Then add 2.5% glutaraldehyde solution to the washed bacteria to make bacteria suspension. Fix at 4°C for 5h, centrifuge at 4000r / min; 10min, use PBS solution to suspend and disperse the bacteria collected by centrifugation to make a suspension. Then let the bacteria sink and dry freely, and sputter gold under vacuum; observe the bacterial structure through a scanning electron microscope, and the working voltage is 10kV. The sample preparation method of the normal bacteria control group was the same as that of the antimicrobial peptide group.
[0034] T...
Embodiment 3
[0037] Embodiment 3: the stability of antibacterial peptide AP22 under different pH value conditions
[0038] Prepare PBS buffer respectively, adjust its pH range to 1-12, take 20 μL of antimicrobial peptide supernatant, and add it to equal volumes of different pH buffers, and use different pH buffers without antimicrobial peptides as a control , Escherichia coli O1 strain was used as the experimental object to carry out the antibacterial experiment, and the antibacterial effect change curve was drawn.
[0039] The results show that the antimicrobial peptide AP22 has certain activity in the pH range of the experiment, and the activity is the largest at the pH value of 5.6-7.4 ( image 3 ).
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