Improvement of Enterocin P Purification Process

S. CUOZZO, S. CALVEZ, H. PREVOST, D. DRIDER

FOLIA MICROBIOLOGICA

Institute of Microbiology, Academy of Sciences of the Czech Republic and Czechoslovak Society for Microbiology

Lugar: Prague; Año: 2006 vol. 51 p. 401 - 405

0015-5632

Purification and heterologous expression of enterocin P (EntP), a sec-dependent bacteriocin produced by Enterococcus faecium, in Escherichia coli is described. PCR-amplified product of the enterocin P structural gene entP was cloned into plasmid pET-32b under the control of the inducible T7lac 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 of E. coli harboring appropriate recombinant plasmid, characterized by ELISA and Western-blot analysis and purified by immunoaffinity chromatography. The use of E. coli as heterologous host and pET-32b as expressing vector offers promising tools for heterologous production of class IIa bacteriocin.