Enterocin P has been characterized in class IIa bacteriocins and showed a broad spectrum of antimicrobial activity. In this study, the nucleotide sequence of enterocin P transformed and expressed in Pichia pastoris genomically, and the antimicrobial activity of the extracted protein was evaluated on pathogenic bacteria. The synthetic sequence of Enterocin P was ligated into pPICZαA, an integrated vector, in the EcoRI - XbaI site. The recombinant plasmid was amplified in Escherichia coli strain DH5α and proved for ligation and probably mutation by sequencing. The linearized plasmid was electroporated into Pichia pastoris. The presence of recombinant protein in yeast cell extraction was verified by SDS-PAGE and the antimicrobial activity of the purified protein. Recombinant Enterocin P tested for the inhibition of 8 avian pathogen bacteria via MIC test. Electrophoresis of purified protein and cell culture supernatant showed a protein band of about 5.4 kDa. The mass spectrometry of the sample exhibits an Amino acid sequence that corresponds to enterocin P in Enterococcus faecium with an accession number of OTN91526.1. Recombinant bacteriocin MIC test shows the most inhibition on Salmonella typhimurium and E. coli O157. Recombinant Pichia pastoris exhibited a potential for producing enterocin P in high concentrations; therefore, this antimicrobial peptide can be studied for pharmacological use or industrial applications.

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