Phosphorylation of glucose by a guanosine-5'-triphosphate (GTP)-dependent glucokinase in Fibrobacter succinogenes subsp. succinogenes S85.
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 pI of 4.82. Double-reciprocal plots of substrate concentration versus velocity were linear and the enzyme had apparent Km values of 55 microM for glucose and 72 microM 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.
Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo 58105.
T L
TL
Glass
J S
JS
Sherwood
D000818:Animals; D002412:Cations; D002417:Cattle; D005941:Glucokinase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D006089:Gram-Negative Anaerobic Bacteria / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D012417:Rumen / Q000382:microbiology; D013379:Substrate Specificity
D002412:Cations; D010710:Phosphates; D006160:Guanosine Triphosphate; D005941:Glucokinase; D005947:Glucose