Cloning and characterization of the biosynthetic gene cluster for kutznerides.
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.
Fujimori
Danica Galonic
DG
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Hrvatin
Sinisa
S
Neumann
Christopher S
CS
Strieker
Matthias
M
Marahiel
Mohamed A
MA
Walsh
Christopher T
CT
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
D047630:Depsipeptides; D000249:Adenosine Monophosphate