Abstract
Kluyveromyces marxianus is thermotolerant yeast that is able to utilize a wider range of substrates and has greater thermal tolerance than most other yeast species. K. marxianus can assimilate xylose, but its ability to produce ethanol from xylose in oxygen-limited environments is poor. In the present study, the K. marxianus xylose reductase (KmXR) gene (Kmxyl1) was cloned and the recombinant enzyme was characterized to clarify the factors that limit xylose fermentation in K. marxianus NBRC1777. KmXR is a key enzyme in the xylose metabolism of K. marxianus, which was verified by disruption of the Kmxyl1 gene. The Km of the recombinant KmXR for NADPH is 65.67 μM and KmXR activity is 1.295 U/mg, which is lower than those of most reported yeast XRs, and the enzyme has no activity with coenzyme NADH. This result demonstrates that the XR from K. marxianus is highly coenzyme specific; combined with the extremely low XDH activity of K. marxianus with NADP+, the limitation of xylose fermentation is due to a redox imbalance under anaerobic conditions and low KmXR activity.
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Acknowledgments
We thank Professor Tamaki Hisanori from Kagoshima University and Professor Kumagai Hidehiko from Ishikawa Prefectural University for providing the K. marxianus YHJ010 and plasmid YEGAP. This work was supported by a grant-in-aid from the National Natural Science Foundation of China (Grant no. 31070028) and the project was also sponsored by National Basic Research Program of China (Grant No. 2011CBA00801), the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry, and the Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant no. 20093402120027).
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Zhang, B., Zhang, L., Wang, D. et al. Identification of a xylose reductase gene in the xylose metabolic pathway of Kluyveromyces marxianus NBRC1777. J Ind Microbiol Biotechnol 38, 2001–2010 (2011). https://doi.org/10.1007/s10295-011-0990-z
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DOI: https://doi.org/10.1007/s10295-011-0990-z