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Characterization of a Recombinant Thermostable Dehalogenase Isolated from the Hot Spring Thermophile Sulfolobus tokodaii

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Abstract

A putative dehalogenase, l-HADST, from the thermophile Sulfolobus tokodaii, was cloned and expressed in Escherichia coli. The recombinant enzyme catalyzes the stereospecific dehalogenation of l-2-haloacids with similar levels of activity as its homolog from mesophiles. l-HADST remains fully active after being incubated for 4 h at 70 °C and tolerates extreme pH conditions ranging from 4 to 10. Furthermore, it can be purified conveniently without the usage of any chromatography method. The high expression yield and easy purification procedure make the recombinant dehalogenase an excellent candidate for biotechnological applications.

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Acknowledgment

This work was supported by the National Institute of Environmental Health Sciences (NIH, grant P42 ES07380). We acknowledge the use of the circular dichroism spectrometer at the Protein Core facility housed in the Department of Molecular and Cellular Biochemistry, University of Kentucky and directed by Dr. David Rodgers.

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Authors and Affiliations

  1. Department of Chemistry, University of Kentucky, Lexington, KY, 40506, USA

    Stacy A. Law & Yinan Wei

  2. Paul Laurence Dunbar High School, 1600 Man O’ War Blvd, Lexington, KY, 40513, USA

    Philip G Bachas-Daunert

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  1. Philip G Bachas-Daunert
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  2. Stacy A. Law
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  3. Yinan Wei
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Correspondence to Yinan Wei.

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Bachas-Daunert, P.G., Law, S.A. & Wei, Y. Characterization of a Recombinant Thermostable Dehalogenase Isolated from the Hot Spring Thermophile Sulfolobus tokodaii . Appl Biochem Biotechnol 159, 382–393 (2009). https://doi.org/10.1007/s12010-009-8589-9

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