title: Two oligopeptide-permease-encoding genes in the clavulanic acid cluster of Streptomyces clavuligerus are essential for production of the beta-lactamase inhibitor.
abstract: orf7 (oppA1) and orf15 (oppA2) are located 8 kb apart in the clavulanic acid gene cluster of Streptomyces clavuligerus and encode proteins which are 48.0% identical. These proteins show sequence similarity to periplasmic oligopeptide-binding proteins. Mutant S. clavuligerus oppA1::acc, disrupted in oppA1, lacks clavulanic acid production. Clavulanic acid production is restored by transformation with plasmid pIJ699-oppA1, which carries oppA1, but not with the multicopy plasmid pIJ699-oppA2, which carries oppA2. The mutant S. clavuligerus oppA2::aph also lacks clavulanic acid production, shows a bald phenotype, and overproduces holomycin (5). Clavulanic acid production at low levels is restored in the oppA2-disrupted mutants by transformation with plasmid pIJ699-oppA2, but it is not complemented by the multicopy plasmid pIJ699-oppA1. Both genes encode oligopeptide permeases with different substrate specificities. The disrupted S. clavuligerus oppA2::aph is not able to grow on RPPGFSPFR (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg; bradykinin), but both mutants grow on VAPG (Val-Ala-Pro-Gly) as the only nitrogen source, indicating differences in the peptide bound by the proteins encoded by both genes. The null S. clavuligerus oppA1::acc and S. clavuligerus oppA2::aph mutants are more resistant to the toxic tripeptide phosphinothricyl-alanyl-alanine (also named bialaphos) than the wild-type strain, suggesting that this peptide might be transported by these peptide-binding proteins.
authors.affiliation: Area de Microbiología, Facultad de Ciencias Biológicas y Ambientales, University of León, 24071 León, Spain.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Lorenzana
authors.forename: Rosario
authors.initials: R
authors.lastname: Pérez-Redondo
authors.forename: Irene
authors.initials: I
authors.lastname: Santamarta
authors.forename: Juan F
authors.initials: JF
authors.lastname: Martín
authors.forename: Paloma
authors.initials: P
authors.lastname: Liras
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D019818:Clavulanic Acid / Q000096:biosynthesis*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family; D009842:Oligopeptides / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics*; D014169:Transformation, Bacterial; D065093:beta-Lactamase Inhibitors
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D026901:Membrane Transport Proteins; D009842:Oligopeptides; D065093:beta-Lactamase Inhibitors; D019818:Clavulanic Acid; C037230:oligopeptide permease, Bacteria