Heterologous extracellular production of enterocin P from Enterococcus faecium P13 in the methylotrophic bacterium Methylobacterium extorquens

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 struct...

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Bibliographic Details
Published in:FEMS microbiology letters 2005-07, Vol.248 (1), p.125-131
Main Authors: Gutiérrez, Jorge, Bourque, Denis, Criado, Raquel, Choi, Young J., Cintas, Luis M., Hernández, Pablo E., Míguez, Carlos B.
Format: Article
Language:English
Subjects:
Antibodies
Bacteria
Bacteriocins
Bacteriocins - biosynthesis
Bacteriocins - genetics
Bacteriology
Biological and medical sciences
Enterocin P
Enterococcus faecium
Enterococcus faecium - genetics
Enterococcus faecium - metabolism
Enzyme-linked immunosorbent assay
Fundamental and applied biological sciences. Psychology
General secretory pathway
Genetics
Heterologous gene expression
Hydrophobicity
Liquid chromatography
Methanol dehydrogenase
Methylobacterium extorquens
Methylobacterium extorquens - genetics
Methylobacterium extorquens - growth & development
Methylobacterium extorquens - metabolism
Microbiology
Pediocin
Plasmids
Purification
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
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Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1016/j.femsle.2005.05.029