title: Improvement of enterocin P purification process.
abstract: 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.
authors.affiliation: Laboratoire de Microbiologie Alimentaire et Industrielle, Ecole Nationale des Ingénieurs des Industries des Techniques Agricoles et Alimentaires, BP 82225 Nantes, France.
authors.forename: S
authors.initials: S
authors.lastname: Cuozzo
authors.forename: S
authors.initials: S
authors.lastname: Calvez
authors.forename: H
authors.initials: H
authors.lastname: Prévost
authors.forename: D
authors.initials: D
authors.lastname: Drider
mesh_terms: D001430:Bacteriocins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D003001:Cloning, Molecular; D016984:Enterococcus faecium / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D007218:Industrial Microbiology / Q000379:methods; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001430:Bacteriocins; D011994:Recombinant Proteins; C417717:enterocin P