Cloning and Analysis of a Type II Polyketide Synthase Gene Cluster from Streptomyces toxytricini NRRL 15,443

  • Yoo An-Na (Faculty of Pharmacy, Yeungnam University) ;
  • Demirev Atanas V. (Faculty of Biotechnology, Yeungnam University) ;
  • Lee, Ji-Seon (Faculty of Biotechnology, Yeungnam University) ;
  • Kim, Sang-Dal (Faculty of Biotechnology, Yeungnam University) ;
  • Nam Doo-Hyun (Faculty of Pharmacy, Yeungnam University)
  • Published : 2006.12.31

Abstract

A standard type II polyketide synthase (PKS) gene cluster was isolated while attempting to clone the biosynthetic gene for lipstatin from Streptomyces toxytricini NRRL 15,443. This result was observed using a Southern blot of a PstI-digested S. toxytricini chromosomal DNA library with a 444 bp amplified probe of a ketosynthase (KS) gene fragment. Four open reading frames [thioesterase (TE), $\beta$-ketoacyl systhase (KAS), chain length factor (CLF), and acyl carrier protein (ACP)], were identified through the nucleotide sequence determination and analysis of a 4.5 kb cloned DNA fragment. In order to confirm the involvement of a cloned gene in lipstatin biosynthesis, a gene disruption experiment for the KS gene was performed. However, the resulting gene disruptant did not show any significant difference in lipstatin production when compared to wild-type S. toxytricini. This result suggests that lipstatin may not be synthesized by a type II PKS.

Keywords

References

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