Improved antimicrobial activities of synthetic-hybrid bacteriocins designed from enterocin E50-52 and pediocin PA-1.

Two hybrid bacteriocins, enterocin E50-52/pediocin PA-1 (EP) and pediocin PA-1/enterocin E50-52 (PE), were designed by combining the N terminus of enterocin E50-52 and the C terminus of pediocin PA-1 and by combining the C terminus of pediocin PA-1 and the N terminus of enterocin E50-52, respectively. Both hybrid bacteriocins showed reduced MICs compared to those of their natural counterparts. The MICs of hybrid PE and EP were 64- and 32-fold lower, respectively, than the MIC of pediocin PA-1 and 8- and 4-fold lower, respectively, than the MIC of enterocin E50-52. In this study, the effect of hybrid as well as wild-type (WT) bacteriocins on the transmembrane electrical potential (ΔΨ) and their ability to induce the efflux of intracellular ATP were investigated. Enterocin E50-52, pediocin PA-1, and hybrid bacteriocin PE were able to dissipate ΔΨ, but EP was unable to deplete this component. Both hybrid bacteriocins caused a loss of the intracellular concentration of ATP. EP, however, caused a faster efflux than PE and enterocin E50-52. Enterocin E50-52 and hybrids PE and EP were active against the Gram-positive and Gram-negative bacteria tested, such as Micrococcus luteus, Salmonella enterica serovar Enteritidis 20E1090, and Escherichia coli O157:H7. The hybrid bacteriocins designed and described herein are antimicrobial peptides with MICs lower those of their natural counterparts. Both hybrid peptides induce the loss of intracellular ATP and are capable of inhibiting Gram-negative bacteria, and PE dissipates the electrical potential. In this study, the MIC of hybrid bacteriocin PE decreased 64-fold compared to the MIC of its natural peptide counterpart, pediocin PA-1. Inhibition of Gram-negative pathogens confers an additional advantage for the application of these peptides in therapeutics.

Antibacterial Activity of Subtilosin Alone and Combined with Curcumin, Poly-Lysine and Zinc Lactate Against Listeria monocytogenes Strains.

In this paper, the antibacterial effects of the Bacillus amyloliquefaciens-produced bacteriocin subtilosin, both alone and in combination with curcumin, ε-poly-L-lysine (poly-lysine), or zinc lactate, were examined against Listeria monocytogenes. Results indicated that subtilosin inhibits both of the studied bacterial strains, Scott A (wild-type, nisin sensitive) and NR30 (nisin resistant). However, L. monocytogenes Scott A was more sensitive to subtilosin and pure curcumin. In addition, subtilosin was more active at an acidic pH. Subtilosin in combination with encapsulated curcumin displayed partial synergy against L. monocytogenes ScottA. It also had synergistic activity against both L. monocytogenes Scott A and L. monocytogenes NR30 when combined with zinc lactate. Only an additive effect was observed for subtilosin when combined with non-encapsulated curcumin or poly-lysine against the mentioned strains. Thus, using the combination of subtilosin with curcumin, poly-lysine, or zinc lactate, a lower effective dose can be used to control L. monocytogenes infection. Our findings suggest that subtilosin could be used as alternative bacteriocin to nisin, providing an opportunity to use a novel natural and efficacious biopreservative against L. monocytogenes in food preservation. This is the first report on the effects of the combination of subtilosin with natural antimicrobials on L. monocytogenes.