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

Ryuya Fujii, Atsushi Minami, Tae Tsukagoshi, Natsuko Sato, Takehiko Sahara, Satoru Ohgiya, Katsuya Gomi, Hideaki Oikawa

Research output: Contribution to journalArticlepeer-review

53 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1813-1817
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume75
Issue number9
DOIs
Publication statusPublished - 2011

Keywords

  • Aphidicolin
  • Aspergillus oryzae
  • Diterpene
  • Heterologous expression
  • Total biosynthesis

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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