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Heterologous expression of the Saccharomyces cerevisiae alcohol acetyltransferase genes in Clostridium acetobutylicum and Escherichia coli for the production of isoamyl acetate

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Journal of Industrial Microbiology and Biotechnology

Abstract

Esters are formed by the condensation of acids with alcohols. The esters isoamyl acetate and butyl butyrate are used for food and beverage flavorings. Alcohol acetyltransferase is one enzyme responsible for the production of esters from acetyl-CoA and different alcohol substrates. The genes ATF1 and ATF2, encoding alcohol acetyltransferases from the yeast Saccharomyces cerevisiae have been sequenced and characterized. The production of acids and alcohols in mass quantities by the industrially important Clostridium acetobutylicum makes it a potential organism for exploitation of alcohol acetyltransferase activity. This report focuses on the heterologous expression of the alcohol acetyltransferases in Escherichia coli and C. acetobutylicum. ATF1 and ATF2 were cloned and expressed in E. coli and ATF2 was expressed in C. acetobutylicum. Isoamyl acetate production from the substrate isoamyl alcohol in E. coli and C. acetobutylicum cultures was determined by head-space gas analysis. Alcohol acetyltransferase I produced more than twice as much isoamyl acetate as alcohol acetyltransferase II when expressed from a high-copy expression vector. The effect of substrate levels on ester production was explored in the two bacterial hosts to demonstrate the efficacy of utilizing ATF1and ATF2 in bacteria for ester production.

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Acknowledgements

We thank Michael Gustin for supplying S. cerevisiae genomic DNA and Miles Scotcher for thoughtful discussions. This research was supported by the National Science Foundation grant BES-0001288, and the Robert A. Welch Foundation grants C-1268 and C-1372.

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Author notes

  1. Ke-Xue Huang

    Present address: R and D Discovery, Dow Agrosciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268, USA

Authors and Affiliations

  1. Department of Biochemistry and Cell Biology, Rice University, 6100 S. Main Street, MS140, Houston, TX 77005, USA

    Catherine Emily Horton, Ke-Xue Huang, George N. Bennett & Frederick B. Rudolph

Authors
  1. Catherine Emily Horton
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  2. Ke-Xue Huang
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  3. George N. Bennett
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  4. Frederick B. Rudolph
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Correspondence to Frederick B. Rudolph.

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Horton, C.E., Huang, KX., Bennett, G.N. et al. Heterologous expression of the Saccharomyces cerevisiae alcohol acetyltransferase genes in Clostridium acetobutylicum and Escherichia coli for the production of isoamyl acetate. J IND MICROBIOL BIOTECHNOL 30, 427–432 (2003). https://doi.org/10.1007/s10295-003-0070-0

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  • DOI: https://doi.org/10.1007/s10295-003-0070-0

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