Abstract
Xylose reductase (XR) is one of the key enzymes for bio-ethanol production from lignocellulosic biomass. Intercellular redox imbalance, caused by different coenzyme specificity of XR and Xylitol dehydrogenase (XDH), has been thought to be one of the main factors of xylitol excretion. We previously succeeded by protein engineering to improve the ethanol production by reverse the XDH dependence from NAD+ to NADP+. In this study, we employed protein engineering to construct a novel strictly NADPH dependent XR from Pichia stipitis by site directed mutagenesis, in order to effective recycling of cofactor between XR and XDH, which subsequently reduce xylitol accumulation. Double mutant E223G/S271A showed strictly NADPH dependent with 90% of wild-type activity.
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Acknowledgment
This work was supported by the New Energy and Industrial Technology Development Organization (NEDO), Japan. It was also supported by a Grant-in-Aid for Young Scientists (B) (no. 21760636 to S.W.) and the Global Center of Excellence (GCOE) program for the “Energy Science in the Age of Global Warming,” a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan.
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Khattab, S.M.R. et al. (2011). Construction of a Novel Strictly NADPH-Dependent Pichia stipitis Xylose Reductase by Site-Directed Mutagenesis for Effective Bioethanol Production. In: Yao, T. (eds) Zero-Carbon Energy Kyoto 2010. Green Energy and Technology. Springer, Tokyo. https://doi.org/10.1007/978-4-431-53910-0_14
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DOI: https://doi.org/10.1007/978-4-431-53910-0_14
Publisher Name: Springer, Tokyo
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