The Journal of General and Applied Microbiology
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Directed evolution and expression tuning of geraniol synthase for efficient geraniol production in Escherichia coli
Miki TashiroAkira FujiiShigeko Kawai-NomaKyoichi SaitoDaisuke Umeno
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Volume 63 (2017) Issue 5 Pages 287-295

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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 5ʹ-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.

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© 2017, Applied Microbiology, Molecular and Cellular Biosciences Research Foundation
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