A Simple and Reliable Method for Determination of Optimum PH in Coupled Enzyme Assays | Zendy

Lee Bowman | Zendy; Rachael Motamed | Zendy; Paul Lee | Zendy; Kadijah Aleem | Zendy; Astha S Berawala | Zendy; Katherine Hayden | Zendy; David J. Bzik | Zendy; Debasish Chattopadhyay | Zendy

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A Simple and Reliable Method for Determination of Optimum PH in Coupled Enzyme Assays

Author(s) -

Lee Bowman,

Rachael Motamed,

Paul Lee,

Kadijah Aleem,

Astha S Berawala,

Katherine Hayden,

David J. Bzik,

Debasish Chattopadhyay

Publication year - 2020

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2019-0126

Subject(s) - enzyme , pyruvate kinase , lactate dehydrogenase , chromatography , chemistry , biochemistry , pyruvate dehydrogenase kinase , pyruvate dehydrogenase complex , glycolysis

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.

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