Directed evolution of squalene synthase for dehydrosqualene biosynthesis

Maiko Furubayashi, Ling Li, Akinori Katabami, Kyoichi Saito, Daisuke Umeno

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Squalene synthase (SQS) catalyzes the first step of sterol/hopanoid biosynthesis in various organisms. It has been long recognized that SQSs share a common ancestor with carotenoid synthases, but it is not known how these enzymes selectively produce their own product. In this study, SQSs from yeast, human, and bacteria were independently subjected to directed evolution for the production of the C30 carotenoid backbone, dehydrosqualene. This was accomplished via high-throughput screening with Pantoea ananatis phytoene desaturase, which can selectively convert dehydrosqualene into yellow carotenoid pigments. Genetic analysis of the resultant mutants revealed various mutations that could effectively convert SQS into a "dehydrosqualene synthase." All of these mutations are clustered around the residues that have been proposed to be important for NADPH binding.

Original languageEnglish
Pages (from-to)3375-3381
Number of pages7
JournalFEBS Letters
Volume588
Issue number18
DOIs
Publication statusPublished - 2014 Sep 17
Externally publishedYes

Keywords

  • Carotenoid synthase
  • Directed evolution
  • Phytoene desaturase
  • Squalene synthase

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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