title: Genetic engineering in Streptomyces roseosporus to produce hybrid lipopeptide antibiotics.
abstract: Daptomycin is a lipopeptide antibiotic produced by a nonribosomal peptide synthetase (NRPS) in Streptomyces roseosporus. The holoenzyme is composed of three subunits, encoded by the dptA, dptBC, and dptD genes, each responsible for incorporating particular amino acids into the peptide. We introduced expression plasmids carrying dptD or NRPS genes encoding subunits from two related lipopeptide biosynthetic pathways into a daptomycin nonproducing strain of S. roseosporus harboring a deletion of dptD. All constructs successfully complemented the deletion in trans, generating three peptide cores related to daptomycin. When these were coupled with incomplete methylation of 1 amino acid and natural variation in the lipid side chain, 18 lipopeptides were generated. Substantial amounts of nine of these compounds were readily obtained by fermentation, and all displayed antibacterial activity against gram-positive pathogens.
authors.affiliation: Cubist Pharmaceuticals, Inc., 65 Hayden Avenue, Lexington, Massachusetts 02421, USA.
authors.forename: Vivian
authors.initials: V
authors.lastname: Miao
authors.forename: Marie-Francois
authors.initials: MF
authors.lastname: Coëffet-Le Gal
authors.forename: Kien
authors.initials: K
authors.lastname: Nguyen
authors.forename: Paul
authors.initials: P
authors.lastname: Brian
authors.forename: Julia
authors.initials: J
authors.lastname: Penn
authors.forename: Andrew
authors.initials: A
authors.lastname: Whiting
authors.forename: Jonathan
authors.initials: J
authors.lastname: Steele
authors.forename: David
authors.initials: D
authors.lastname: Kau
authors.forename: Steve
authors.initials: S
authors.lastname: Martin
authors.forename: Robert
authors.initials: R
authors.lastname: Ford
authors.forename: Trevor
authors.initials: T
authors.lastname: Gibson
authors.forename: Mario
authors.initials: M
authors.lastname: Bouchard
authors.forename: Stephen K
authors.initials: SK
authors.lastname: Wrigley
authors.forename: Richard H
authors.initials: RH
authors.lastname: Baltz
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000494:pharmacology*; D001483:Base Sequence; D003001:Cloning, Molecular; D017576:Daptomycin / Q000737:chemistry / Q000494:pharmacology; D005818:Genetic Engineering; D016908:Gram-Positive Bacterial Infections / Q000188:drug therapy; D008074:Lipoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology*; D008745:Methylation; D008969:Molecular Sequence Data; D005810:Multigene Family; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D021122:Protein Subunits; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D008074:Lipoproteins; D021122:Protein Subunits; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; D017576:Daptomycin