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Engineering redox cofactor utilization for detoxification of glycolaldehyde, a key inhibitor of bioethanol production, in yeast Saccharomyces cerevisiae

  • Bioenergy and biofuels
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Applied Microbiology and Biotechnology Aims and scope Submit manuscript

An Erratum to this article was published on 21 June 2014

Abstract

Hot-compressed water treatment of lignocellulose liberates numerous inhibitors that prevent ethanol fermentation of yeast Saccharomyces cerevisiae. Glycolaldehyde is one of the strongest fermentation inhibitors and we developed a tolerant strain by overexpressing ADH1 encoding an NADH-dependent reductase; however, its recovery was partial. In this study, to overcome this technical barrier, redox cofactor preference of glycolaldehyde detoxification was investigated. Glycolaldehyde-reducing activity of the ADH1-overexpressing strain was NADH-dependent but not NADPH-dependent. Moreover, genes encoding components of the pentose phosphate pathway, which generates intracellular NADPH, was upregulated in response to high concentrations of glycolaldehyde. Mutants defective in pentose phosphate pathways were sensitive to glycolaldehyde. Genome-wide survey identified GRE2 encoding a NADPH-dependent reductase as the gene that confers tolerance to glycolaldehyde. Overexpression of GRE2 in addition to ADH1 further improved the tolerance to glycolaldehyde. NADPH-dependent glycolaldehyde conversion to ethylene glycol and NADP+ content of the strain overexpressing both ADH1 and GRE2 were increased at 5 mM glycolaldehyde. Expression of GRE2 was increased in response to glycolaldehyde. Carbon metabolism of the strain was rerouted from glycerol to ethanol. Thus, it was concluded that the overexpression of GRE2 together with ADH1 restores glycolaldehyde tolerance by augmenting the NADPH-dependent reduction pathway in addition to NADH-dependent reduction pathway. The redox cofactor control for detoxification of glycolaldehyde proposed in this study could influence strategies for improving the tolerance of other fermentation inhibitors.

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Acknowledgments

We would like to acknowledge the Analytical Research Center for Experimental Sciences of Saga University for DNA sequencing. Part of this study was supported by JSPS KAKENHI (24580117) to HK.

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Authors and Affiliations

  1. Department of Environmental Science, Faculty of Agriculture, Saga University, Saga, 8408502, Japan

    Lahiru N. Jayakody, Kenta Horie, Nobuyuki Hayashi & Hiroshi Kitagaki

  2. Department of Biochemistry and Applied Biosciences, United Graduate School of Agricultural Sciences Kagoshima, Kagoshima University, Korimoto 1-21-24, Kagoshima, 890-8580, Japan

    Lahiru N. Jayakody, Nobuyuki Hayashi & Hiroshi Kitagaki

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  1. Lahiru N. Jayakody
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  2. Kenta Horie
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Correspondence to Hiroshi Kitagaki.

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Jayakody, L.N., Horie, K., Hayashi, N. et al. Engineering redox cofactor utilization for detoxification of glycolaldehyde, a key inhibitor of bioethanol production, in yeast Saccharomyces cerevisiae . Appl Microbiol Biotechnol 97, 6589–6600 (2013). https://doi.org/10.1007/s00253-013-4997-4

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  • DOI: https://doi.org/10.1007/s00253-013-4997-4

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