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Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase
Published on: December 19, 2011
Assaying for pyruvate, orthophosphate dikinase activity: Necessary precautions with phosphoenolpyruvate carboxylase
G Salahas1, Y Manetas, N A Gavalas
1Laboratory of Plant Physiology, Department of Biology, University of Patras, Patras, Greece.
Abstract:
Phosphoenolpyruvate carboxylase (EC 4.1.1.31), used as a coupling enzyme in the assay of the pyruvate, orthophosphate dikinase (EC 2.7.9.1) forward reaction, is a serious limiting factor for the overall rate when added at a level of 0.2-0.3 unit/ml of assay medium. Nonlimiting assay conditions are obtained by either increasing the level of the coupling enzyme to 3 units/ml or adding 6mM glucose-6-phosphate as an activator/stabilizer of phosphoenolpyruvate carboxylase.
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