The antibacterial activity and mechanism analysis of piscidin 5 like from Larimichthys crocea

Chemical drugs, such as antibiotics, were still important materials to prevent and cure diseases of aquatic organisms. However, antibiotics abuse do not only make the effects little, but also cause other bad problems, such as bacterial resistance and drug residues. Therefore, seeking the effective substitutes of antibiotics was an approach needed to be explored. Antibacterial peptides (AMPs) attracted more and more attention in the recent years. The parasitism and secondary bacterial invasion caused by ectroparasite Cryptocaryon irritans was a disaster to almost all host fish, including Larimichthys crocea. Reports indicated many AMPs played a key role in the whole parasitic infection cycle. Piscidin 5 like was a member of piscidin family. In the study, the antibacterial activity and mechanisms of piscidin 5 like from L.coreca (Lc-P5L) were detected. Liquid growth inhibition results showed recombinant Lc-P5L (rLc-P5L) had broad antibacterial spectrum and strong bactericidal activity. The bactericidal activity functioned in dose- and time-dependent manners. SEM (scanning electron microscope) observed the relatively detailed bactericidal process, rLc-P5L treatment resulted in a mass of bacteria piling together, appearing plenty of strange filaments and covering on the bacteria. Besides, S.aureus overgrowed plenty of granules, formed holes on the membrane of a few cells, and contents poured out from the holes. At the same time, antibacterial mechanisms were explored. After direct incubation with bacteria, western blot detected the apparently positive signal of rLc-P5L on bacteria; secondly, the incubation first with LPS (lipopolysaccharide) or LTA (lipoteichoic acid) significantly affect the binding of rLc-P5L to bacteria again, which indicated rLc-P5L could bind to bacteria through interaction with some PAMPs (pathogen-associated molecular patterns). In addition, rLc-P5L could interact with bacterial genome DNA by dose- and time-dependent means. In summary, rLc-P5L binded to bacteria surface through targeting to some PAMPs to damage membrane, and entered into cells to interact with genome DNA to disturb normal metabolism when it reached to some certain time and concentration thresholds, which were likely to be its pathway to exert antibacterial activity.