Abstract:In order to establish the application platform for antibiotic and antiseptic application of Lactobacillus rhamnosus bacteriocin, the author explored its antibacterial properties. The main regulatory gene of Lactobacillus rhamnosus was found to be wo-a/2, which was determined by DNASTAR as the whole gene sequence analysis software. The wo-a/2 gene was expressed by prokaryotic expression method and its solubility, induced expression conditions, immunogenicity, hemolytic activity and antibacterial characteristics were analyzed. The results showed that pET-32a-wo-a/2 plasmid was successfully constructed and expressed Lactobacillus rhamnosus bacteriocin WO-A/2, which mainly existed in the form of inclusion body. When pET-32a-wo-a/2 plasmid was inoculated in LB liquid medium containing Amp+ and induced with IPTG at a final concentration of 1.0 mmol/L for 8 h, the expression of bacteriocin WO-A/2 reached the maximum. It was verified from the double-layer Oxford cup blood agar plate method that Lactobacillus rhamnosus bacteriocin WO-A/2 had no hemolytic characteristics either at different concentrations. Bacteriocin WO-A/2 was found to promote the apoptosis of Staphylococcus aureus, as observed by dual fluorescence staining and laser confocal microscopy. This study provides a reference for Lactobacillus rhamnosus bacterin to be a safe and reliable antibiotic agent.
