Directed evolution and expression tuning of geraniol synthase for efficient geraniol production in escherichia coli

Miki Tashiro, Akira Fujii, Shigeko Kawai-Noma, Kyoichi Saito, Daisuke Umeno

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

To achieve an efficient production of geraniol and its derivatives in Escherichia coli, we aimed to improve the activity of geraniol synthase (GES) through a single round of mutagenesis and screening for higher substrate consumption. We isolated GES variants that outperform their parent in geraniol production. The analysis of GES variants indicated that the expression level of GES was the bottleneck for geraniol synthesis. Over-expression of the mutant GESM53 with a 5′-untranslated sequence designed for high translational efficiency, along with the additional expression of mevalonate pathway enzymes, isopentenyl pyrophosphate isomerase, and geranyl pyrophosphate synthase, yielded 300 mg/L/12 h geraniol and its derivatives (>1000 mg/L/42 h in total) in a shaking flask.

Original languageEnglish
Pages (from-to)287-295
Number of pages9
JournalJournal of General and Applied Microbiology
Volume63
Issue number5
DOIs
Publication statusPublished - 2017
Externally publishedYes

Keywords

  • High-throughput screening
  • Metabolic engineering
  • Mevalonate pathway
  • Monoterpene
  • RBS score

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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