title: Cloning and characterization of the biosynthetic gene cluster for kutznerides.
abstract: Kutznerides, actinomycete-derived cyclic depsipetides, consist of six nonproteinogenic residues, including a highly oxygenated tricyclic hexahydropyrroloindole, a chlorinated piperazic acid, 2-(1-methylcyclopropyl)-glycine, a beta-branched-hydroxy acid, and 3-hydroxy glutamic acid, for which biosynthetic logic has not been elucidated. Herein we describe the biosynthetic gene cluster for the kutzneride family, identified by degenerate primer PCR for halogenating enzymes postulated to be involved in biosyntheses of these unusual monomers. The 56-kb gene cluster encodes a series of six nonribosomal peptide synthetase (NRPS) modules distributed over three proteins and a variety of tailoring enzymes, including both mononuclear nonheme iron and two flavin-dependent halogenases, and an array of oxygen transfer catalysts. The sequence and organization of NRPS genes support incorporation of the unusual monomer units into the densely functionalized scaffold of kutznerides. Our work provides insight into the formation of this intriguing class of compounds and provides a foundation for elucidating the timing and mechanisms of their biosynthesis.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Danica Galonic
authors.initials: DG
authors.lastname: Fujimori
authors.forename: Sinisa
authors.initials: S
authors.lastname: Hrvatin
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Neumann
authors.forename: Matthias
authors.initials: M
authors.lastname: Strieker
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Marahiel
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Walsh
mesh_terms: D000192:Actinomycetales / Q000235:genetics* / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D047630:Depsipeptides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family
chemical_list: D047630:Depsipeptides; D000249:Adenosine Monophosphate