title: An asparagine oxygenase (AsnO) and a 3-hydroxyasparaginyl phosphotransferase (HasP) are involved in the biosynthesis of calcium-dependent lipopeptide antibiotics.
abstract: Nonribosomal peptides contain a wide range of unusual non-proteinogenic amino acid residues. As a result, these complex natural products are amongst the most structurally diverse secondary metabolites in nature, and possess a broad spectrum of biological activities. beta-Hydroxylation of amino acid precursors or peptidyl residues and their subsequent processing by downstream tailoring enzymes are some of the most common themes in the biosynthetic diversification of these therapeutically important peptides. Identification and characterization of the biosynthetic intermediates and enzymes involved in these processes are thus pivotal in understanding nonribosomal peptide assembly and modification. To this end, the putative asparaginyl oxygenase- and 3-hydroxyasparaginyl phosphotransferase-encoding genes hasP and asnO were separately deleted from the calcium-dependent antibiotic (CDA) biosynthetic gene cluster of Streptomyces coelicolor. Whilst the parent strains produce a number of 3-hydroxyasparagine- and 3-phosphohydroxyasparagine-containing CDAs, the DeltahasP mutants produce exclusively non-phosphorylated CDAs. On the other hand, DeltaasnO mutants produce several new Asn-containing CDAs not present in the wild-type, which retain calcium-dependent antimicrobial activity. This confirms that AsnO and HasP are required for the beta-hydroxylation and phosphorylation of the Asn residue within CDA.
authors.affiliation: Department of Biomolecular Sciences, UMIST, PO Box 88, Manchester M60 1QD, UK.|School of Chemistry, The University of Manchester, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Joanne M
authors.initials: JM
authors.lastname: Neary
authors.affiliation: School of Chemistry, The University of Manchester, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Amanda
authors.initials: A
authors.lastname: Powell
authors.affiliation: Department of Biomolecular Sciences, UMIST, PO Box 88, Manchester M60 1QD, UK.|School of Chemistry, The University of Manchester, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Lyndsey
authors.initials: L
authors.lastname: Gordon
authors.affiliation: School of Chemistry, The University of Manchester, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Claire
authors.initials: C
authors.lastname: Milne
authors.affiliation: Department of Biomolecular Sciences, UMIST, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Fiona
authors.initials: F
authors.lastname: Flett
authors.affiliation: Biotica Technology Ltd, Chesterford Research Park, Little Chesterford, Saffron Walden, Essex CB10 1XL, UK.
authors.forename: Barrie
authors.initials: B
authors.lastname: Wilkinson
authors.affiliation: Department of Biomolecular Sciences, UMIST, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Colin P
authors.initials: CP
authors.lastname: Smith
authors.affiliation: School of Chemistry, The University of Manchester, PO Box 88, Manchester M60 1QD, UK.
authors.forename: Jason
authors.initials: J
authors.lastname: Micklefield
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000898:Antibiosis; D001216:Asparagine / Q000378:metabolism*; D001407:Bacillus / Q000187:drug effects; D017353:Gene Deletion; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D016982:Micrococcus luteus / Q000187:drug effects; D015394:Molecular Structure; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D046908:Peptide Biosynthesis, Nucleic Acid-Independent; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism*; D048372:Streptomyces coelicolor / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; C040487:CDA antibiotic; D010455:Peptides; D001216:Asparagine; D010105:Oxygenases; D010770:Phosphotransferases