Expression of the hybrid bacteriocin Ent35-MccV in Saccharomyces cerevisiae

LANZA, L; ACUÑA, L; MINAHK, C.J; BELLOMIO, A; CHALON M.C.

Cordoba

Congreso; Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular; 2016

Sociedad Argentina de Investigaciones en Bioquimica y Biologia Molecular

The hybrid bacteriocin Ent35-MccV is a construction made by the fusion of genes encoding Enterocin CRL35 (munA) and Microcin V (cvaC). Enterocin CRL35 is active against the foodborne pathogen Listeria monocytogenes. On the other hand, Microcin V is active specifically against Gram-negative bacteria. These bacteriocins were fused in order to obtain a peptide with broad antimicrobial spectrum. The aim of this work was to express Ent35-MccV in Saccharomyces cerevisiae, a GRAS microorganism, and evaluate its activity against Gram-positive and negative pathogen bacteria. To achieve this, munA-cvaC with a leader peptide were cloned into the yeast-E. coli shuttle plasmid pCEV-G1. S. cerevisiae 23344C was transformed by the lithium acetate method. The expression of the hybrid Ent35-MccV was induced with the addition of galactose. The hybrid bacteriocin Ent35-MccV was obtained from the yeast producing supernatants. The antibiotic activity was assayed against different pathogen bacteria. The chimerical bacteriocin displayed antimicrobial activity against E. coli 0157, L. monocytogenes and Salmonella gallinarum biovar gallinarum and pullorum, strains that infects chickens. Therefore, the hybrid bacteriocin may find important applications in veterinary to control salmonellosis in poultry.