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Overexpression in a non-native halophilic host and biotechnological potential of NAD+-dependent glutamate dehydrogenase from Halobacterium salinarum strain NRC-36014

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Abstract

Enzymes produced by halophilic archaea are generally heat resistant and organic solvent tolerant, and accordingly important for biocatalytic applications in ‘green chemistry’, frequently requiring a low-water environment. NAD+-dependent glutamate dehydrogenase from an extremely halophilic archaeon Halobacterium salinarum strain NRC-36014 was selected to explore the biotechnological potential of this enzyme and genetically engineered derivatives. Over-expression in a halophilic host Haloferax volcanii provided a soluble, active recombinant enzyme, not achievable in mesophilic Escherichia coli, and an efficient purification procedure was developed. pH and salt dependence, thermostability, organic solvent stability and kinetic parameters were explored. The enzyme is active up to 90 °C and fully stable up to 70 °C. It shows good tolerance of various miscible organic solvents. High concentrations of salt may be substituted with 30 % DMSO or betaine with good stability and activity. The robustness of this enzyme under a wide range of conditions offers a promising scaffold for protein engineering.

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Acknowledgments

N. M. was supported by an Ad Astra scholarship from University College Dublin. We are also grateful to Dr. Peter Lund and Dr. Andrew Large (University of Birmingham) for providing us the halophilic expression vector and host strain and to Science Foundation Ireland for a Research Fellowship (2006–2011) to P.C.E.

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Authors and Affiliations

  1. Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin 4, Ireland

    Nayla Munawar

  2. UCD School of Biomolecular and Biomedical Sciences, Conway Institute, University College Dublin, Belfield, Dublin 4, Republic of Ireland

    Paul C. Engel

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  1. Nayla Munawar
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  2. Paul C. Engel
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Correspondence to Paul C. Engel.

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Communicated by F. Robb.

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Munawar, N., Engel, P.C. Overexpression in a non-native halophilic host and biotechnological potential of NAD+-dependent glutamate dehydrogenase from Halobacterium salinarum strain NRC-36014. Extremophiles 16, 463–476 (2012). https://doi.org/10.1007/s00792-012-0446-z

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