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°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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August 2002
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Research Article|
August 15 2002
Biochemical adaptations of two sugar kinases from the hyperthermophilic archaeon Pyrococcus furiosus
Corné H. VERHEES;
Corné H. VERHEES
1
∗Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, The Netherlands
1To whom correspondence should be addressed (e-mail corne.verhees@algemeen.micr.wau.nl).
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Denise G.M. KOOT;
Denise G.M. KOOT
∗Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, The Netherlands
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Thijs J.G. ETTEMA;
Thijs J.G. ETTEMA
∗Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, The Netherlands
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Cor DIJKEMA;
Cor DIJKEMA
†Laboratory of Biophysics, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands
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Willem M. de VOS;
Willem M. de VOS
∗Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, The Netherlands
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John van der OOST
John van der OOST
∗Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, The Netherlands
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Publisher: Portland Press Ltd
Received:
October 30 2001
Revision Received:
April 17 2002
Accepted:
April 29 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 366 (1): 121–127.
Article history
Received:
October 30 2001
Revision Received:
April 17 2002
Accepted:
April 29 2002
Citation
Corné H. VERHEES, Denise G.M. KOOT, Thijs J.G. ETTEMA, Cor DIJKEMA, Willem M. de VOS, John van der OOST; Biochemical adaptations of two sugar kinases from the hyperthermophilic archaeon Pyrococcus furiosus. Biochem J 15 August 2002; 366 (1): 121–127. doi: https://doi.org/10.1042/bj20011597
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