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Identification of a xylulokinase catalyzing xylulose phosphorylation in the xylose metabolic pathway of Kluyveromyces marxianus NBRC1777

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Xylulokinase is one of the key enzymes in xylose metabolism and fermentation, and fine-tuned expression of xylulokinase can improve xylose fermentation in yeast. To improve the efficiency of xylose fermentation in Kluyveromyces marxianus, the gene KmXYL3, which encodes a d-xylulokinase (E.C. 2.7.1.17), was isolated from K. marxianus NBRC1777. KmXYL3 was expressed in Escherichia coli BL21 (DE3) cells, and the specific activity of the resulting recombinant purified xylulokinase was 23.5 mU/mg. Disruption of KmXYL3 resulted in both loss of xylitol utilization and marked decrease in xylose utilization, proving that KmXYL3 encodes a xylulokinase that catalyzes the reaction from xylulose to xylulose 5-phosphate in the xylose metabolic pathway. The slow assimilation of xylose observed in the KmXYL3-disrupted strain indicates that KmXYL3 is critical for xylose and xylitol utilization; however, K. marxianus utilizes a bypass pathway for xylose assimilation, and this pathway does not involve xylitol or xylulose.

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Acknowledgments

We thank Professor Tamaki Hisanori from Kagoshima University and Kumagai Hidehiko from Ishikawa Prefectural University for providing us 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 the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry and Specialized Research Fund for the Doctoral Program of Higher Education of China (grant no. 20093402120027) and the Fundamental Research Funds for the Central Universities (grant no. WK2070000007).

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Authors and Affiliations

  1. School of Life Science, University of Science and Technology of China, Hefei, Anhui, 230027, People’s Republic of China

    Rongliang Wang, Dongmei Wang, Xiaolian Gao & Jiong Hong

  2. School of Life Science and Engineering, Southwest University of Science and Technology, Mianyang, Sichuan, 621010, People’s Republic of China

    Ling Zhang

  3. Department of Biology and Biochemistry, University of Houston, Houston, TX, 77004-5001, USA

    Xiaolian Gao

  4. Hefei National Laboratory for Physical Science at the Microscale, Hefei, Anhui, 230026, People’s Republic of China

    Rongliang Wang, Xiaolian Gao & Jiong Hong

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  1. Rongliang Wang
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  2. Ling Zhang
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  3. Dongmei Wang
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Correspondence to Jiong Hong.

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Wang, R., Zhang, L., Wang, D. et al. Identification of a xylulokinase catalyzing xylulose phosphorylation in the xylose metabolic pathway of Kluyveromyces marxianus NBRC1777. J Ind Microbiol Biotechnol 38, 1739–1746 (2011). https://doi.org/10.1007/s10295-011-0963-2

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  • DOI: https://doi.org/10.1007/s10295-011-0963-2

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