Abstract
Cell extracts of Fibrobacter succinogenes subsp. succinogenes S85 phosphorylated glucose with a GTP-dependent glucokinase. The enzyme showed little activity with ATP (12% of that with GTP). Of other phosphate donors tested, only dGTP and ITP gave high glucokinase activities. Dialyzed extracts required Mg+2 and K+ for maximal activity. In potassium phosphate buffer, glucokinase showed maximum activity at pH 7.5 with glucose-6-phosphate dehydrogenase as the coupling enzyme. In this assay, glucokinase was active with glucose (100%), 2-deoxy-d-glucose (40%), and mannose (20%). Partially purified glucokinase had a molecular weight of 82,000 and a pl of 4.82. Double-reciprocal plots of substrate concentration versus velocity were linear and the enzyme had apparent Km values of 55 μM for glucose and 72 μM for GTP. Dialyzed cell extracts of Fibrobacter intestinalis C1A also contained a GTP-dependent glucokinase that showed little activity with ATP. Potassium also stimulated the activity of this enzyme. These results suggest that this unusual glucokinase may be characteristic of the genus Fibrobacter.
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Abbreviations
- CHES :
-
cyclohexylaminoethanesulfonic acid
- GK :
-
glucokinase
- PEP :
-
phosphoenolpyruvate
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Published with the approval of the Director of the North Dakota Agricultural Experiment Station as journal article no. 2186
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Glass, T.L., Sherwood, J.S. Phosphorylation of glucose by a guanosine-5′-triphosphate (GTP)-dependent glucokinase in Fibrobacter succinogenes subsp. succinogenes S85. Arch Microbiol 162, 180–186 (1994). https://doi.org/10.1007/BF00314472
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DOI: https://doi.org/10.1007/BF00314472