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Discovery and characterization of a novel ATP/polyphosphate xylulokinase from a hyperthermophilic bacterium Thermotoga maritima

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Xylulokinase (XK, E.C. 2.7.1.17) is one of the key enzymes in xylose metabolism and it is essential for the activation of pentoses for the sustainable production of biocommodities from biomass sugars. The open reading frame (TM0116) from the hyperthermophilic bacterium Thermotoga maritima MSB8 encoding a putative xylulokinase were cloned and expressed in Escherichia coli BL21 Star (DE3) in the Luria–Bertani and auto-inducing high-cell-density media. The basic biochemical properties of this thermophilic XK were characterized. This XK has the optimal temperature of 85 °C. Under a suboptimal condition of 60 °C, the k cat was 83 s−1, and the K m values for xylulose and ATP were 1.24 and 0.71 mM, respectively. We hypothesized that this XK could work on polyphosphate possibly because this ancestral thermophilic microorganism utilizes polyphosphate to regulate the Embden–Meyerhof pathway and its substrate-binding residues are somewhat similar to those of other ATP/polyphosphate-dependent kinases. This XK was found to work on low-cost polyphosphate, exhibiting 41 % of its specific activity on ATP. This first ATP/polyphosphate XK could have a great potential for xylose utilization in thermophilic ethanol-producing microorganisms and cell-free biosystems for low-cost biomanufacturing without the use of ATP.

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Acknowledgments

This work was supported by the Biological Systems Engineering Department of Virginia Tech, and partially supported Shell by GameChanger Program, the CALS Biodesign and Bioprocessing Research Center, and NSF SBIR grant to PZ. JSMC thanks the Mexican Council of Science and Technology (Conacyt) for her PhD study.

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Authors and Affiliations

  1. Biological Systems Engineering Department, Virginia Tech, 304 Seitz Hall, Blacksburg, VA, 24061, USA

    Julia S. Martín del Campo, You Chun, Jae-Eung Kim & Y.-H. Percival Zhang

  2. Departamento de Física Aplicada, Centro de Investigación y de Estudios Avanzados–Unidad Mérida, Carretera antigua a Progreso Km. 6, A.P. 73 Cordemex, 97310, Mérida, Yucatán, Mexico

    Julia S. Martín del Campo & Rodrigo Patiño

  3. Institute for Critical Technology and Applied Sciences (ICTAS), Virginia Tech, Blacksburg, VA, 24061, USA

    You Chun & Y.-H. Percival Zhang

  4. Cell Free Bioinnovations Inc., Blacksburg, VA, 24060, USA

    Y.-H. Percival Zhang

  5. Gate Fuels Inc., Blacksburg, VA, 24060, USA

    Y.-H. Percival Zhang

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  1. Julia S. Martín del Campo
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  2. You Chun
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  3. Jae-Eung Kim
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  5. Y.-H. Percival Zhang
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Correspondence to Y.-H. Percival Zhang.

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Martín del Campo, J.S., Chun, Y., Kim, JE. et al. Discovery and characterization of a novel ATP/polyphosphate xylulokinase from a hyperthermophilic bacterium Thermotoga maritima . J Ind Microbiol Biotechnol 40, 661–669 (2013). https://doi.org/10.1007/s10295-013-1265-7

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