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Expanding ester biosynthesis in Escherichia coli

Nature Chemical Biology volume 10, pages 259–265 (2014)Cite this article

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Abstract

To expand the capabilities of whole-cell biocatalysis, we have engineered Escherichia coli to produce various esters. The alcohol O-acyltransferase (ATF) class of enzyme uses acyl-CoA units for ester formation. The release of free CoA upon esterification with an alcohol provides the free energy to facilitate ester formation. The diversity of CoA molecules found in nature in combination with various alcohol biosynthetic pathways allows for the biosynthesis of a multitude of esters. Small to medium volatile esters have extensive applications in the flavor, fragrance, cosmetic, solvent, paint and coating industries. The present work enables the production of these compounds by designing several ester pathways in E. coli. The engineered pathways generated acetate esters of ethyl, propyl, isobutyl, 2-methyl-1-butyl, 3-methyl-1-butyl and 2-phenylethyl alcohols. In particular, we achieved high-level production of isobutyl acetate from glucose (17.2 g l−1). This strategy was expanded to realize pathways for tetradecyl acetate and several isobutyrate esters.

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Figure 1: Enzymatic ester synthesis by combining alcohols and acyl-CoAs.
Figure 2: Constructing acetate ester synthesis pathways in E. coli.
Figure 3: Tetradecyl acetate production from glucose in E. coli.
Figure 4: Constructing higher chain ester biosynthesis pathway in E. coli.

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Acknowledgements

This work was supported by University of California–Davis startup fund and the Hellman fellowship to S.A. G.M.R. is supported by a US National Institutes of Health Biotechnology Training Grant Fellowship (T32-GM008799) and a Sloan Fellowship. Y.T. is supported by Japan Society for the Promotion of Science postdoctoral fellowship for research abroad. We would like to thank R. Luu and R.E. Parales (University of California–Davis) for graciously providing genomic DNA from P. putida g7 and M. Kato and S.-J. Lin (University of California–Davis) for providing genomic DNA from S. cerevisiae BY4742. We also thank M.D. Toney and C.A. Rabinovitch-Deere (University of California–Davis) for critical reading of the manuscript. Finally, we thank S. Desai for technical assistance with HPLC analysis and for constructing pAL603.

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  1. Gabriel M Rodriguez and Yohei Tashiro: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of California–Davis, Davis, California, USA

    Gabriel M Rodriguez, Yohei Tashiro & Shota Atsumi

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  1. Gabriel M Rodriguez
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Contributions

G.M.R., Y.T. and S.A. designed research; G.M.R. and Y.T. performed the experiments; G.M.R., Y.T. and S.A. analyzed data; and G.M.R., Y.T. and S.A. wrote the paper.

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Correspondence to Shota Atsumi.

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Rodriguez, G., Tashiro, Y. & Atsumi, S. Expanding ester biosynthesis in Escherichia coli. Nat Chem Biol 10, 259–265 (2014). https://doi.org/10.1038/nchembio.1476

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