Improvement of enterocin P purification process

Abstract

Purification and heterologous expression of enterocin P (EntP), a sec-dependent bacteriocin produced byEnterococcus faecium, inEscherichia coli is described. PCR-amplified product of the enterocin P structural geneentP was cloned into plasmid pET-32b under the control of the inducible T7/ac promoter. The neo-synthesized EntP was genetically modified by an addition of 3 extra amino acids, leading to recombinant EntRP. Active EntRP was recovered from the cytoplasmic soluble fraction ofE. coli harboring appropriate recombinant plasmid, characterized by ELISA and Western-blot analysis and purified by immunoaffinity chromatography. The use ofE. coli as heterologous host and pET-32b as expressing vector offers promising tools for heterologous production of class IIa bacteriocin.