title: Heterologous Processing and Export of the Bacteriocins Pediocin PA-1 and Lactococcin A in Lactococcus Lactis: A Study with Leader Exchange.
abstract: The bacteriocins pediocin PA-1 and lactococcin A are synthesized as precursors carrying N-terminal extensions with a conserved cleavage site preceded by two glycine residues in positions -2 and -1. Each bacteriocin is translocated through the cytoplasmic membrane by an integral membrane protein of the ABC cassette superfamily which, in the case of pediocin PA-1, has been shown to possess peptidase activity responsible for proteolytic cleavage of the pre-bacteriocin. In each case, another integral membrane protein is essential for bacteriocin production. In this study, a two-step PCR approach was used to permutate the leaders of pediocin PA-1 and lactococcin A. Wild-type and chimeric pre-bacteriocins were assayed for maturation by the processing/export machinery of pediocin PA-1 and lactococcin A. The results show that pediocin PA-1 can be efficiently exported by the lactococcin machinery whether it carries the lactococcin or the pediocin leader. It can also compete with wild-type lactococcin A for the lactococcin machinery. Pediocin PA-1 carrying the lactococcin A leader or lactococcin A carrying that of pediocin PA-1 was poorly secreted when complemented with the pediocin PA-1 machinery, showing that the pediocin machinery is more specific for its bacteriocin substrate. Wild-type pre-pediocin and chimeric pre-pediocin were shown to be processed by the lactococcin machinery at or near the double-glycine cleavage site. These results show the potential of the lactococcin LcnC/LcnD machinery as a maturation system for peptides carrying double-glycine-type amino-terminal leaders.
authors.affiliation: Department of Food Science, Rutgers, The State University of New Jersey, 65 Dudley Road, New Brunswick, NJ, 08901-8520, USA. tchikindas@aesop.rutgers.edu.
authors.forename: M
authors.initials: M
authors.lastname: Chikindas
authors.affiliation: Département des sciences des aliments et de nutrition, Centre de recherche STELA, Université Laval, Quebec, QC, G1K 7P4, Canada.
authors.forename: E
authors.initials: E
authors.lastname: Emond
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands.
authors.forename: A J
authors.initials: AJ
authors.lastname: Haandrikman
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Kok
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Leenhouts
authors.affiliation: Département des sciences des aliments et de nutrition, Centre de recherche STELA, Université Laval, Quebec, QC, G1K 7P4, Canada.
authors.forename: S
authors.initials: S
authors.lastname: Pandian
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands.
authors.forename: G
authors.initials: G
authors.lastname: Venema
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Venema
keywords: Bacteriocin; Double-glycine leader; Lactococcin; Pediocin