Cloning of the gene cluster responsible for the biosynthesis of brasilicardin A, a unique diterpenoid

J Antibiot (Tokyo). 2008 Mar;61(3):164-74. doi: 10.1038/ja.2008.126.

Abstract

Brasilicardin A (BCA), produced by Nocardia brasiliensis IFM 0406 (currently referred to as N. terpenica), has a unique structure consisting of a diterpene skeleton with L-rhamnose, N-acetylglucosamine, amino acid, and 3-hydroxybenzoate moieties, and exhibits potent biological activities. To understand the biosynthetic machinery of this unique compound, we have cloned the corresponding gene cluster. Firstly, we cloned a gene by PCR that encodes geranylgeranyl diphosphate synthase (GGPPS), which produces a direct precursor of diterpene compounds. We obtained four candidate genes and one of the genes was confirmed to encode a GGPPS. By sequence analysis of regions flanking the GGPPS gene, we identified eleven genes (bra1-11), all oriented in the same direction. We did not, however, detect any genes related to L-rhamnose and N-acetylglucosamine biosyntheses in the flanking regions. A gene disruption experiment did indeed show that this gene cluster was responsible for BCA biosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aminoglycosides / biosynthesis*
  • Aminoglycosides / genetics*
  • Cloning, Molecular
  • Diterpenes / chemistry*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Molecular Sequence Data
  • Multigene Family
  • Nocardia / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Aminoglycosides
  • Diterpenes
  • brasilicardin A