Open AccessHeterologous extracellular production of enterocin P from Enterococcus faecium P13 in the methylotrophic bacterium Methylobacterium extorquens
Author(s) -
Gutiérrez Jorge,
Bourque Denis,
Criado Raquel,
Choi Young J.,
Cintas Luis M.,
Hernández Pablo E.,
Míguez Carlos B.
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.05.029
Subject(s) - enterococcus faecium , methanol dehydrogenase , bacteriocin , plasmid , heterologous expression , heterologous , biology , recombinant dna , bacteria , microbiology and biotechnology , methylobacterium , biochemistry , gene , genetics , 16s ribosomal rna
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.