Investigation of the Antibacterial Effectiveness of Hybrid Recombinant Bacteriocins Circular Enterocin and Pyocin L1 Derived from Pseudomonas aeruginosa and Enterococcus faecalis.

Abstract

This study investigates the antibacterial effectiveness of hybrid recombinant bacteriocins, specifically circular enterocin and pyocin L1, derived from Pseudomonas aeruginosa and Enterococcus faecalis. Novel hybrid variations, such as H-PyCir, were produced by combining, expressing, and isolating the genes responsible for these bacteriocins. The hybrid bacteriocins were purified and tested against a wide range of bacterial strains, demonstrating strong antibacterial activity against both Gram-positive and Gram-negative bacteria. The results reveal that hybrid bacteriocins, particularly H-PyCir, exhibit significantly enhanced antibacterial efficacy compared to their individual parent bacteriocins. Techniques such as phylogenetic analysis, plasmid construction, 3D homology modeling, and in silico docking were employed to investigate the structural and functional properties of these hybrids, providing insights into their mechanisms of action. The study also conducted well diffusion assays, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) tests, which confirmed the superior antimicrobial activity of H-PyCir, with the lowest MIC and MBC values observed across all tested strains. SDS-PAGE analysis further validated the successful cloning, expression, and purification of the hybrid bacteriocins, indicating high purity and correct molecular weights. These results highlight the potential of hybrid bacteriocins as promising alternatives to traditional antibiotics, offering a novel therapeutic approach to combat antibiotic resistance.