To explore the potential intracellular mechanism of the antimicrobial peptide HJH-3 in killing Salmonella, a DNA blocking test and scanning electron microscopy (SEM) were used to determine the ability of the peptide to bind bacterial DNA in vitro. Laser confocal analysis and electron microscopy were used to observe the binding of antimicrobial peptide HJH-3 and Salmonella DNA, and flow cytometry was used to analyze the effect of antimicrobial peptides on cell division in vivo. The results showed that HJH-3 can bind to DNA to block the diffusion and migration of DNA in agarose gel. Laser confocal microscopy revealed that antimicrobial peptide HJH-3 penetrated the bacterial cell membrane and bound with bacterial DNA. Transmission electron microscopy showed that antimicrobial peptide HJH-3 aggregated in the nucleoid of Salmonella cells, and through a channel in the membrane destroyed by the antimicrobial peptide, DNA and other intracellular contents were excreted, and polymerized DNA was fragmented. The results of the flow cytometry analysis confirmed that the death rate of Salmonella increased significantly after exposure to antimicrobial peptide HJH-3 and increased with increasing antimicrobial peptide concentration. These results suggest that AMP HJH-3 may be a candidate antimicrobial agent to treat infectious diseases caused by Salmonella pullorum.
This journal is © The Royal Society of Chemistry.