Journal
JOURNAL OF BIOTECHNOLOGY
Volume 165, Issue 3-4, Pages 153-156Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2013.03.009
Keywords
Xylose fermentation; Xylose reductase; Xylitol dehydrogenase; Coenzyme specificity; Ethanol production
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Funding
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- Global Center of Excellence (GCOE) program for the Energy Science in the Age of Global Warming
- Ministry of Education, Culture, Sports, Science and Technology in Japan [23603003]
- Iwatani Foundation
- Grants-in-Aid for Scientific Research [23603003] Funding Source: KAKEN
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The xylose-fermenting recombinant Saccharomyces cerevisiae and its improvement have been studied extensively. The redox balance between xylose reductase (XR) and xylitol dehydrogenase (XDH) is thought to be an important factor in effective xylose fermentation. Using protein engineering, we previously successfully reduced xylitol accumulation and improved ethanol production by reversing the dependency of XDH from NAD(+) to NADP(+). We also constructed a set of novel strictly NADPH-dependent XR from Pichia stipitis by site-directed mutagenesis. In the present study, we constructed a set of recombinant S. cerevisiae carrying a novel set of mutated strictly NADPH-dependent XR and NADP(+)-dependent XDH genes with overexpression of endogenous xylulokinase (XK) to study the effects of complete NADPH/NADP(+) recycling on ethanol fermentation and xylitol accumulation. All mutated strains demonstrated reduced xylitol accumulation, ranging 34.4-54.7% compared with the control strain. Moreover, compared with the control strain, the two strains showed 20% and 10% improvement in ethanol production. (C) 2013 Elsevier B.V. All rights reserved.
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