Characterization of a bacteriocin produced by enterococcus gallinarum CRL 1826 Isolated from captive bullfrog: evaluation of its mode of action against Listeria monocytogenes and gram-negatives

Abstract

Enterococcus gallinarum CRL 1826 isolated from an American bullfrog (Lithobates catesbeianus) skin inhibits the growth of Citrobacter freundii, Pseudomonas aeruginosa (bullfrog pathogens) and Listeria monocytogenes by a synergistic effect between organic acids and a bacteriocin-like molecule. This bacteriocin, named enterocin CRL 1826, showed a proteinaceous nature, heat stability and polar characteristics. Its production followed kinetics of primary metabolites synthesis reaching a maximum of 61,400 AU/mL. The minimum inhibitory and minimumbactericidal concentrations were 2,640 and 5,280 AU/mL, respectively, against L. monocytogenes. The addition of 120,000 AU/mL of enterocin to growing L. monocytogenes and Gram-negative (P. aeruginosa and C. freundii) bacteria showed bactericidal and bacteriostatic effects, respectively. However, enterocin derived-peptides had bactericidal effect only against Gram-negatives.Enterocin produced cell envelope damages and efflux of citosolic content on L. monocytogenes, while enterocin derived-peptides showed granulation and contraction of cytoplasm material on P. aeruginosa and increase in theperiplasmic space and empty cells appearance on C. freundii.Enterocin CRL 1826 is the first bacteriocin described for E. gallinarum from raniculture. It could be used as a biopreservative while the derived-peptides represent an alternative to control multi-drug resistant Gram-negatives.The antimicrobial spectrum and the stability of enterocin and its derived-peptides indicate that they could be applied in different biotechnological areas.