title: Heterologous extracellular production of enterocin P from Enterococcus faecium P13 in the methylotrophic bacterium Methylobacterium extorquens.
abstract: Enterocin P (EntP), a strong antilisterial pediocin-like bacteriocin from Enterococcus faecium P13, was produced by Methylobacterium extorquens. For heterologous expression of EntP in the methylotrophic bacterium M. extorquens, a recombinant plasmid was constructed. The gene encoding the EntP structural gene (entP) was cloned into the plasmid vector pCM80, under control of the methanol dehydrogenase promoter (P(mxaF)), to generate plasmid pS25. When M. extorquens ATCC 55366 was transformed with pS25, EntP was detected and quantified in supernatants of the recombinant M. extorquens S25 strain by using specific anti-EntP antibodies and a non-competitive indirect enzyme-linked immunosorbent assay (NCI-ELISA). Purification of EntP by hydrophobic adsorption and reverse-phase (RP-FPLC) chromatographies, permitted recovery of active EntP from the supernatants of M. extorquens S25 grown in a synthetic defined medium.
authors.affiliation: Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Gutiérrez
authors.forename: Denis
authors.initials: D
authors.lastname: Bourque
authors.forename: Raquel
authors.initials: R
authors.lastname: Criado
authors.forename: Young J
authors.initials: YJ
authors.lastname: Choi
authors.forename: Luis M
authors.initials: LM
authors.lastname: Cintas
authors.forename: Pablo E
authors.initials: PE
authors.lastname: Hernández
authors.forename: Carlos B
authors.initials: CB
authors.lastname: Míguez
mesh_terms: D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D016984:Enterococcus faecium / Q000235:genetics* / Q000378:metabolism; D020601:Methylobacterium extorquens / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
chemical_list: D001430:Bacteriocins; D011994:Recombinant Proteins; C417717:enterocin P