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. 2002 Aug 15;366(Pt 1):121–127. doi: 10.1042/BJ20011597

Biochemical adaptations of two sugar kinases from the hyperthermophilic archaeon Pyrococcus furiosus.

Corné H Verhees 1, Denise G M Koot 1, Thijs J G Ettema 1, Cor Dijkema 1, Willem M de Vos 1, John van der Oost 1
PMCID: PMC1222741  PMID: 11978175

Abstract

The hyperthermophilic archaeon Pyrococcus furiosus possesses a modified Embden-Meyerhof pathway, including an unusual ADP-dependent glucokinase (ADP-GLK) and an ADP-dependent phosphofructokinase. In the present study, we report the characterization of a P. furiosus galactokinase (GALK) and its comparison with the P. furiosus ADP-GLK. The pyrococcal genes encoding the ADP-GLK and GALK were functionally expressed in Escherichia coli, and the proteins were subsequently purified to homogeneity. Both enzymes are specific kinases with an optimal activity at approx. 90 degrees C. Biochemical characterization of these enzymes confirmed that the ADP-GLK is unable to use ATP as the phosphoryl group donor, but revealed that GALK is ATP-dependent and has an extremely high affinity for ATP. There is a discussion about whether the unusual features of these two classes of kinases might reflect adaptations to a relatively low intracellular ATP concentration in the hyperthermophilic archaeon P. furiosus.

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