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A highly thermostable trehalase from the thermophilic bacterium Rhodothermus marinus

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Abstract

Trehalases play a central role in the metabolism of trehalose and can be found in a wide variety of organisms. A periplasmic trehalase (α,α-trehalose glucohydrolase, EC 3.2.1.28) from the thermophilic bacterium Rhodothermus marinus was purified and the respective encoding gene was identified, cloned and overexpressed in Escherichia coli. The recombinant trehalase is a monomeric protein with a molecular mass of 59 kDa. Maximum activity was observed at 88°C and pH 6.5. The recombinant trehalase exhibited a K m of 0.16 mM and a V max of 81 μmol of trehalose (min)−1 (mg of protein)−1 at the optimal temperature for growth of R. marinus (65°C) and pH 6.5. The enzyme was highly specific for trehalose and was inhibited by glucose with a K i of 7 mM. This is the most thermostable trehalase ever characterized. Moreover, this is the first report on the identification and characterization of a trehalase from a thermophilic bacterium.

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Acknowledgments

This work was funded by the European Commission Contracts QLK3-CT-2000-00640 and COOP-CT-2003-508644 and Fundação para a Ciência e a Tecnologia and FEDER, Portugal, POCI/59310/2004. We thank Winfried Boos, Konstanz, for a fruitful collaboration that led to the discovery of trehalase activity in R. marinus. M. Regalla from Analytical Services performed the N-terminal sequencing at the Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal. C. Jorge acknowledges a PhD grant from PRAXIS XXI (SFRH/BD/10572/2002).

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Authors and Affiliations

  1. Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Rua da Quinta Grande 6, Apartado 127, 2780-156, Oeiras, Portugal

    Carla D. Jorge, Maria Manuel Sampaio & Helena Santos

  2. Prokaria Ltd, Gylfaflot 5, 112 Reykjavik, Iceland

    Gudmundur Ó. Hreggvidsson & Jakob K. Kristjánson

  3. University of Iceland, Sudurgata, 101 Reykjavik, Iceland

    Gudmundur Ó. Hreggvidsson

Authors
  1. Carla D. Jorge
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  2. Maria Manuel Sampaio
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  3. Gudmundur Ó. Hreggvidsson
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  4. Jakob K. Kristjánson
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  5. Helena Santos
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Corresponding author

Correspondence to Helena Santos.

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Communicated by G. Antranikian

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Jorge, C.D., Sampaio, M.M., Hreggvidsson, G.Ó. et al. A highly thermostable trehalase from the thermophilic bacterium Rhodothermus marinus . Extremophiles 11, 115–122 (2007). https://doi.org/10.1007/s00792-006-0021-6

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  • DOI: https://doi.org/10.1007/s00792-006-0021-6

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