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Activation of Pyrococcus furiosus alkaline phosphatase by divalent metal ions

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Abstract

Treatment of a hyperthermophilic enzyme, alkaline phosphatase from Pyrococcus furiosus (PfuAP), with EDTA completely deactivated PfuAP, indicating that the presence of one or more divalent metal ions is essential for its catalytic activity. Subsequent addition of various divalent metal ions to the apoprotein recovered the enzymatic activity and, in particular, the addition of Co(II) resulted in an over 50-fold increase in activity compared with PfuAP before EDTA treatment. Intriguingly, PfuAP with Co(II) exhibited weaker stability toward heat treatment, suggesting that Co2+ destabilizes the tertiary structure of PfuAP at high temperature.

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Acknowledgments

This work was supported in part by the Science and Technology Incubation Program in Advanced Regions from the Japan Science and Technology Agency (JST). K. Minamihata was supported by a Research Fellowship of the Japan Society for the Promotion of Science (JSPS) for young scientists.

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Authors and Affiliations

  1. Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka, 819-0395, Japan

    Kosuke Minamihata, Masahiro Goto & Noriho Kamiya

  2. Center for Future Chemistry, Kyushu University, 744 Motooka, Fukuoka, 819-0395, Japan

    Masahiro Goto & Noriho Kamiya

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  1. Kosuke Minamihata
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  2. Masahiro Goto
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  3. Noriho Kamiya
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Correspondence to Noriho Kamiya.

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Minamihata, K., Goto, M. & Kamiya, N. Activation of Pyrococcus furiosus alkaline phosphatase by divalent metal ions. Biotechnol Lett 34, 2055–2060 (2012). https://doi.org/10.1007/s10529-012-0998-0

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  • DOI: https://doi.org/10.1007/s10529-012-0998-0

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