The development of new antimicrobial peptides has become an attractive alternative to conventional antibiotics due to the increasing rates of microbial drug resistance. Ib-M corresponds to a family of cationic synthetic peptides, 20 amino acids in length, that have shown inhibitory effect against the non-pathogenic strain Escherichia coli K-12. This work evaluated the antimicrobial potential of Ib-M peptides against the pathogenic E. coli O157: H7 using a reference strain and a clinical isolate. The Ib-M peptides showed antibacterial activity against both strains of E. coli O157: H7; the minimum inhibitory concentration of Ib-M peptides ranged from 1.6 to 12.5 μM and the minimum bactericidal concentration ranged from 3.7 to 22.9 μM, being Ib-M1 and Ib-M2 the peptides that presented the highest inhibitory effect. Time-kill kinetics assay showed a reduction of the bacterial population by more than 95% after 4 hours of exposure to 1xMIC of Ib-M1. Low cytotoxicity was observed in VERO cells with 50% cytotoxic concentration in the range from 197.5 to more than 400 μM. All peptides showed a random structure in hydrophilic environments, except Ib-M1, and all of them transitioned to an α-helical structure when the hydrophobicity of the medium was increased. In conclusion, these findings support the in vitro antimicrobial effect of Ib-M peptides against the pathogenic bacteria E. coli O157: H7 and prove to be promising molecules for the development of new therapeutic alternatives.

The Ib-M1 peptide exhibits bactericidal activity against Escherichia coli O157:H7 and low toxicity in mammalian cells. The present study aimed to evaluate the effect of Ib-M1 on E. coli O157:H7 membrane permeabilization. For this purpose, the minimum inhibitory concentrations of Ib-M1 for E. coli O157:H7 and ML35 were measured. The permeability of the E. coli outer and inner membranes was determined by measuring N-phenyl-1-naphthylamine and O-nitrophenyl-β-galactosidase hydrolysis, respectively after bacterial exposure to antimicrobial peptides and control antibiotics. Morphological changes in antimicrobial-exposed E. coli O157:H7 were evaluated by scanning electron microscopy following treatment with antimicrobial peptides. Ib-M1 expressed activity against E. coli O157:H7 and ML35 at minimal inhibitory concentrations (MIC) of 2.9 ± 1.7 and 6.3 ± 0 μM, respectively. The peptide induced permeabilization of the outer membrane of E. coli O157:H7 at all concentrations evaluated and permeabilization of the inner membrane after 50 min at concentrations between 1× MIC and 8× MIC. The morphological changes induced by Ib-M1 led to significant alterations in bacterial shape including collapsed cells and pronounced surface roughness and invaginations. In conclusion, physiological and morphological evidence indicates that the Ib-M1 antimicrobial effect against E. coli O157:H7 is mediated by its permeabilizing action on the outer and inner bacterial membranes.