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A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
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Growth-phase dependent expression of the mevalonate pathway in a terpenoid antibiotic-producing Streptomyces strain

Yoshimitsu Hamano1, Tohru Dairi, Masahiro Yamamoto

  • 1Biotechnology Research Center, Toyama Prefectural University, Japan.

Bioscience, Biotechnology, and Biochemistry
|May 31, 2002
PubMed

Insights

Streptomyces griseolosporeus uses two pathways for IPP production. The nonmevalonate pathway supports primary metabolism, while the mevalonate pathway is crucial for terpentecin antibiotic production during secondary metabolism.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • Streptomyces griseolosporeus MF730-N6 produces the terpenoid antibiotic terpentecin (Tp).
  • This strain possesses both nonmevalonate and mevalonate pathways for isopentenyl pyrophosphate (IPP) synthesis.
  • Previous work cloned the Tp biosynthetic gene (ter) and the mevalonate pathway gene cluster (mev).

Purpose of the Study:

  • To clone and characterize genes involved in the nonmevalonate pathway (dxs 1, dxs 2, dxr) in S. griseolosporeus.
  • To investigate the temporal expression of nonmevalonate and mevalonate pathway genes in relation to Tp production.
  • To elucidate the roles of these pathways in primary and secondary metabolism.

Main Methods:

  • Cloning of dxs 1, dxs 2, dxr, and gap genes.
  • Expression of cloned genes in E. coli to confirm enzymatic activity.
  • Northern blot and primer extension analyses to determine gene expression patterns.
  • Quantitative analysis of gene transcripts throughout cultivation.

Main Results:

  • Two distinct dxs genes (dxs 1, dxs 2) and a dxr gene from the nonmevalonate pathway were successfully cloned and their activities confirmed.
  • Transcripts for dxs 1, dxs 2, dxr, and the housekeeping gap gene were detected throughout the cultivation period.
  • Mevalonate (mev) and terpentecin (ter) pathway genes were not expressed during the early growth phase but were upregulated upon initiation of Tp production.

Conclusions:

  • The nonmevalonate pathway is primarily involved in primary metabolism, while the mevalonate pathway is predominantly utilized for secondary metabolism (Tp production).
  • Both dxs genes are actively transcribed in Streptomyces griseolosporeus.
  • This study provides insights into the coordinated regulation of primary and secondary metabolic pathways in antibiotic production.

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