Total biosynthesis of diterpene aphidicolin, a specific inhibitor of DNA polymerase α: heterologous expression of four biosynthetic genes in Aspergillus oryzae

Biosci Biotechnol Biochem. 2011;75(9):1813-7. doi: 10.1271/bbb.110366. Epub 2011 Sep 7.


Clustering of biosynthetic genes for producing fungal secondary metabolites, which frequently consist of less than ten genes, has been recognized with numerous genomes. The heterologous expression of whole genes in the clusters will therefore produce various types of natural products when using a suitable fungal host. We introduced the whole gene cluster for the biosynthesis of diterpene aphidicolin into the fungal quadruple auxotrophic host, Aspergillus oryzae, by using four different vectors (pTAex3, pPTRI, pUSA and pAdeA) which harbor a starch-inducible promoter/terminator to examine the expression conditions. The resulting quadruple transformant carrying the genes of geranylgeranyl diphosphate synthase PbGGS, terpene synthase PbACS, and two monooxygenases (PbP450-1 and PbP450-2) produced aphidicolin. The double and triple transformants also respectively produced aphidicolan-16β-ol and 3-deoxyaphidicolin. Alternative host Saccharomyces cerevisiae carrying the genes, PbGGS and PbACS, produced key intermediate aphidicolan-16β-ol. This is the first example of a total biosynthesis of terpenoids using fungal hosts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkyl and Aryl Transferases / genetics
  • Alkyl and Aryl Transferases / metabolism*
  • Aphidicolin / biosynthesis*
  • Aspergillus oryzae / enzymology
  • Aspergillus oryzae / genetics*
  • DNA Polymerase I / antagonists & inhibitors
  • Diterpenes / chemistry
  • Diterpenes / metabolism
  • Enzyme Inhibitors / metabolism*
  • Enzyme Inhibitors / pharmacology
  • Farnesyltranstransferase / genetics
  • Farnesyltranstransferase / metabolism*
  • Gas Chromatography-Mass Spectrometry
  • Genes, Fungal
  • Genetic Engineering / methods
  • Genetic Vectors
  • Mixed Function Oxygenases / genetics
  • Mixed Function Oxygenases / metabolism*
  • Multigene Family
  • Promoter Regions, Genetic
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Terpenes / chemistry
  • Terpenes / metabolism*
  • Transformation, Genetic


  • Diterpenes
  • Enzyme Inhibitors
  • Terpenes
  • Aphidicolin
  • 3-deoxyaphidicolin
  • Mixed Function Oxygenases
  • Alkyl and Aryl Transferases
  • terpene synthase
  • Farnesyltranstransferase
  • DNA Polymerase I