The nonribosomal peptide synthetase enzyme DdaD tethers N(β)-fumaramoyl-l-2,3-diaminopropionate for Fe(II)/α-ketoglutarate-dependent epoxidation by DdaC during dapdiamide antibiotic biosynthesis

J Am Chem Soc. 2010 Nov 10;132(44):15773-81. doi: 10.1021/ja1072367.

Abstract

The gene cluster from Pantoea agglomerans responsible for biosynthesis of the dapdiamide antibiotics encodes an adenylation-thiolation didomain protein, DdaD, and an Fe(II)/α-ketoglutarate-dependent dioxygenase homologue, DdaC. Here we show that DdaD, a nonribosomal peptide synthetase module, activates and sequesters N(β)-fumaramoyl-l-2,3-diaminopropionate as a covalently tethered thioester for subsequent oxidative modification of the fumaramoyl group. DdaC catalyzes Fe(II)- and α-ketoglutarate-dependent epoxidation of the covalently bound N(β)-fumaramoyl-l-2,3-diaminopropionyl-S-DdaD species to generate N(β)-epoxysuccinamoyl-DAP (DAP = 2,3-diaminopropionate) in thioester linkage to DdaD. After hydrolytic release, N(β)-epoxysuccinamoyl-DAP can be ligated to l-valine by the ATP-dependent ligase DdaF to form the natural antibiotic N(β)-epoxysuccinamoyl-DAP-Val.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Anti-Bacterial Agents / biosynthesis*
  • Epoxy Compounds / chemistry*
  • Iron / chemistry*
  • Ketoglutaric Acids / chemistry*
  • Molecular Structure
  • Multigene Family
  • Pantoea / enzymology*
  • Pantoea / genetics
  • Pantoea / metabolism
  • Peptide Synthases / antagonists & inhibitors
  • Peptide Synthases / chemistry*
  • Peptide Synthases / genetics

Substances

  • Anti-Bacterial Agents
  • Epoxy Compounds
  • Ketoglutaric Acids
  • Iron
  • Peptide Synthases