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Studies on the mechanism of synthesis of ethyl acetate in Kluyveromyces marxianus DSM 5422

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Abstract

Kluyveromyces marxianus converts whey-borne sugar into ethyl acetate, an environmentally friendly solvent with many applications. K. marxianus DSM 5422 presumably synthesizes ethyl acetate from acetyl-SCoA. Iron limitation as a trigger for this synthesis is explained by a diminished aconitase and succinate dehydrogenase activity (both enzymes depend on iron) causing diversion of acetyl-SCoA from the tricarboxic acid cycle to ester synthesis. Copper limitation as another trigger for ester synthesis in this yeast refers to involvement of the electron transport chain (all ETC complexes depend on iron and complex IV requires copper). This hypothesis was checked by using several ETC inhibitors. Malonate was ineffective but carboxin partially inhibited complex II and initiated ester synthesis. Antimycin A and cyanide as complexes III and IV inhibitors initiated ester synthesis only at moderate levels while higher concentrations disrupted all respiration and caused ethanol formation. A restricted supply of oxygen (the terminal electron acceptor) also initiated some ester synthesis but primarily forced ethanol production. A switch from aerobic to anaerobic conditions nearly stopped ester synthesis and induced ethanol formation. Iron-limited ester formation was compared with anaerobic ethanol production; the ester yield was lower than the ethanol yield but a higher market price, a reduced number of process stages, a faster process, and decreased expenses for product recovery by stripping favor biotechnological ester production.

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Acknowledgments

Thanet Urit would like to express his thanks to the Rajabhat Nakhon Sawan University (Muang Nakhon Sawan, Thailand) for financial support. We are grateful to Mrs. E. Kneschke for technical assistance, to M. Heller from the Sachsenmilch Leppersdorf GmbH (Germany) for providing whey permeate, and to A. Stößer and A. Stukert for performing some of the presented experiments.

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  1. Institute of Food Technology and Bioprocess Engineering, TU Dresden, 01062, Dresden, Germany

    Christian Löser, Thanet Urit, Peter Keil & Thomas Bley

  2. Department of Biology and Biotechnology, Faculty of Science and Technology, Nakhon Sawan Rajabhat University, Nakhon Sawan, 60000, Thailand

    Thanet Urit

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  1. Christian Löser
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Löser, C., Urit, T., Keil, P. et al. Studies on the mechanism of synthesis of ethyl acetate in Kluyveromyces marxianus DSM 5422. Appl Microbiol Biotechnol 99, 1131–1144 (2015). https://doi.org/10.1007/s00253-014-6098-4

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