A Simple and Reliable Method for Determination of Optimum PH in Coupled Enzyme Assays
Lee Bowman, Rachael Motamed, Paul Lee, Kadijah Aleem, Astha S Berawala, Katherine Hayden, David J. Bzik, Debasish Chattopadhyay
Abstract
Determination of the optimum pH in a coupled enzyme assay poses significant challenges because altering the pH of the reaction mixture can affect the performance of both enzymes. Here, we demonstrate a simple and reliable method to determine the pH optimum for pyruvate kinase using the pyruvate kinase/lactate dehydrogenase coupled enzyme assay. This simple and reliable method can be broadly adapted to determine the pH optimum for various enzymes that are assayed using a coupled enzyme assay.
Topics & Concepts
EnzymePyruvate kinaseLactate dehydrogenaseChromatographyChemistryBiochemistryPyruvate dehydrogenase kinasePyruvate dehydrogenase complexGlycolysisAmino Acid Enzymes and MetabolismMicrobial Metabolic Engineering and BioproductionBiochemical and Molecular Research