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Bifunctional phosphoglucose/phosphomannose isomerase from the hyperthermophilic archaeon Pyrobaculum aerophilum - Extremophiles

  • ️Schönheit, Peter
  • ️Tue Aug 03 2004

Abstract

ORF PAE1610 from the hyperthermophilic crenarchaeon Pyrobaculum aerophilum was first annotated as the conjectural pgi gene coding for hypothetical phosphoglucose isomerase (PGI). However, we have recently identified this ORF as the putative pgi/pmi gene coding for hypothetical bifunctional phosphoglucose/phosphomannose isomerase (PGI/PMI). To prove its coding function, ORF PAE1610 was overexpressed in Escherichia coli, and the recombinant enzyme was characterized. The 65-kDa homodimeric protein catalyzed the isomerization of both glucose-6-phosphate and mannose-6-phosphate to fructose-6-phosphate at similar catalytic rates, thus characterizing the enzyme as bifunctional PGI/PMI. The enzyme was extremely thermoactive; it had a temperature optimum for catalytic activity of about 100°C and a melting temperature for thermal unfolding above 100°C.

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Acknowledgements

We thank B. Siebers, H.P. Klenk, W. Liebl, and P. Redder for providing us—prior to publication—the PGI/PMI sequences from T. tenax, H. butylicus, P. torridus, and S. acidocaldarius, respectively. The expert technical assistance of A. Brandenburger and K. Lutter-Mohr is gratefully acknowledged. Some preliminary experiments on cloning and expression of PaPGI/PMI were performed by D. Knierim in an advanced student course. This work was supported by grants of the Deutsche Forschungsgemeinschaft (SCHO 316/9-1) and the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Institut für Allgemeine Mikrobiologie, Christian-Albrechts-Universität Kiel, Am Botanischen Garten 1-9, 24118, Kiel, Germany

    Thomas Hansen & Peter Schönheit

  2. Institut für Biophysikalisch-Biochemische Verfahren Medizinische Hochschule, Carl Neuberg Str. 1, 30625, Hannover, Germany

    Claus Urbanke

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  1. Thomas Hansen

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  2. Claus Urbanke

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  3. Peter Schönheit

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Correspondence to Peter Schönheit.

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Communicated by G. Antranikian

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Hansen, T., Urbanke, C. & Schönheit, P. Bifunctional phosphoglucose/phosphomannose isomerase from the hyperthermophilic archaeon Pyrobaculum aerophilum. Extremophiles 8, 507–512 (2004). https://doi.org/10.1007/s00792-004-0411-6

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  • Received: 23 March 2004

  • Accepted: 14 June 2004

  • Published: 03 August 2004

  • Issue Date: December 2004

  • DOI: https://doi.org/10.1007/s00792-004-0411-6

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