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Properties of recombinant Strep-tagged and untagged hyperthermophilic D-arabitol dehydrogenase from Thermotoga maritima

  • Biotechnologically Relevant Enzymes and Proteins
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An Erratum to this article was published on 15 April 2011

Abstract

The first hyperthermophilic d-arabitol dehydrogenase from Thermotoga maritima was heterologously purified from Escherichia coli. The protein was purified with and without a Strep-tag. The enzyme exclusively catalyzed the NAD(H)-dependent oxidoreduction of d-arabitol, d-xylitol, d-ribulose, or d-xylulose. A twofold increase of catalytic rates was observed upon addition of Mg2+ or K+. Interestingly, only the tag-less protein was thermostable, retaining 90% of its activity after 90 min at 85 °C. However, the tag-less form of d-arabitol dehydrogenase had similar kinetic parameters compared to the tagged enzyme, demonstrating that the Strep-tag was not deleterious to protein function but decreased protein stability. A single band at 27.6 kDa was observed on SDS-PAGE and native PAGE revealed that the protein formed a homohexamer and a homododecamer. The enzyme catalyzed oxidation of d-arabitol to d-ribulose and therefore belongs to the class of d-arabitol 2-dehydrogenases, which are typically observed in yeast and not bacteria. The product d-ribulose is a rare ketopentose sugar that has numerous industrially applications. Given its thermostability and specificity, d-arabitol 2-dehydrogenase is a desirable biocatalyst for the production of rare sugar precursors.

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Acknowledgments

This project was supported by funds from Bundesministerium für Bildung und Forschung (BMBF, project no. 0315632A).

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Authors and Affiliations

  1. Institut für Mikrobiologie & Biotechnologie, Universität Bonn, 168 Meckenheimer Allee, 53515, Bonn, Germany

    Verena Kallnik, Christian Schultz, Paul Schweiger & Uwe Deppenmeier

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  1. Verena Kallnik
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  2. Christian Schultz
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  3. Paul Schweiger
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  4. Uwe Deppenmeier
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Correspondence to Uwe Deppenmeier.

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An erratum to this article can be found at https://doi.org/10.1007/s00253-011-3250-2

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Kallnik, V., Schultz, C., Schweiger, P. et al. Properties of recombinant Strep-tagged and untagged hyperthermophilic D-arabitol dehydrogenase from Thermotoga maritima . Appl Microbiol Biotechnol 90, 1285–1293 (2011). https://doi.org/10.1007/s00253-011-3187-5

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