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The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) of Sulfolobus solfataricus: a key-enzyme of the semi-phosphorylative branch of the Entner–Doudoroff pathway

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Abstract

Archaea utilize a branched modification of the classical Entner–Doudoroff (ED) pathway for sugar degradation. The semi-phosphorylative branch merges at the level of glyceraldehyde 3-phosphate (GAP) with the lower common shunt of the Emden-Meyerhof-Parnas pathway. In Sulfolobus solfataricus two different GAP converting enzymes—classical phosphorylating GAP dehydrogenase (GAPDH) and the non-phosphorylating GAPDH (GAPN)—were identified. In Sulfolobales the GAPN encoding gene is found adjacent to the ED gene cluster suggesting a function in the regulation of the semi-phosphorylative ED branch. The biochemical characterization of the recombinant GAPN of S. solfataricus revealed that—like the well-characterized GAPN from Thermoproteus tenax—the enzyme of S. solfataricus exhibits allosteric properties. However, both enzymes show some unexpected differences in co-substrate specificity as well as regulatory fine-tuning, which seem to reflect an adaptation to the different lifestyles of both organisms. Phylogenetic analyses and database searches in Archaea indicated a preferred distribution of GAPN (and/or GAP oxidoreductase) in hyperthermophilic Archaea supporting the previously suggested role of GAPN in metabolic thermoadaptation. This work suggests an important role of GAPN in the regulation of carbon degradation via modifications of the EMP and the branched ED pathway in hyperthermophilic Archaea.

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Abbreviations

ED:

Entner–Doudoroff

sp:

Semi-phosphorylative

np:

Non-phosphorylative

EMP:

Embden-Meyerhof-Parnas

G1P:

Glucose 1-phosphate

GAP:

Glyceraldehyde 3-phosphate

GAPN:

Non-phosphorylating GAP dehydrogenase

GAPDH:

GAP dehydrogenase

GAPOR:

GAP oxidoreductase

KDG:

2-Keto-3-deoxygluconate

KDPG:

2-Keto-3-deoxy-6-phosphogluconate

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Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft (SPP 1112, SI642/6-1) and by a Rubicon grant to T.E. from the Netherlands Organisation for Scientific Research (NWO).

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Authors and Affiliations

  1. Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, Norbyvägen 18C, 752 36, Uppsala, Sweden

    Thijs J. G. Ettema

  2. Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands

    Thijs J. G. Ettema, Ans C. M. Geerling & John van der Oost

  3. Department of Biology and Geography, Institute of Biology, Microbiology I, Universität Duisburg-Essen, Universitätsstr. 5, 45117, Essen, Germany

    Hatim Ahmed & Bettina Siebers

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  1. Thijs J. G. Ettema
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  2. Hatim Ahmed
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  3. Ans C. M. Geerling
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  4. John van der Oost
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Correspondence to Bettina Siebers.

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Communicated by D.A. Cowan.

Thijs J. G. Ettema and Hatim Ahmed have contributed equally to this work.

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Ettema, T.J.G., Ahmed, H., Geerling, A.C.M. et al. The non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) of Sulfolobus solfataricus: a key-enzyme of the semi-phosphorylative branch of the Entner–Doudoroff pathway. Extremophiles 12, 75–88 (2008). https://doi.org/10.1007/s00792-007-0082-1

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