A simple and reliable method for determination of optimum pH in coupled enzyme assays

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 kina...

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Main Authors: Lee Bowman, Rachael Motamed, Paul Lee, Kadijah Aleem, Astha S Berawala, Katherine L Hayden, David J Bzik, Debasish Chattopadhyay
Format: Article
Language:English
Published: Taylor & Francis Group 2020-04-01
Series:BioTechniques
Subjects:
Online Access:https://www.future-science.com/doi/10.2144/btn-2019-0126
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author Lee Bowman
Rachael Motamed
Paul Lee
Kadijah Aleem
Astha S Berawala
Katherine L Hayden
David J Bzik
Debasish Chattopadhyay
author_facet Lee Bowman
Rachael Motamed
Paul Lee
Kadijah Aleem
Astha S Berawala
Katherine L Hayden
David J Bzik
Debasish Chattopadhyay
author_sort Lee Bowman
collection DOAJ
description 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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series BioTechniques
spelling doaj-art-bb7edf781d0e48d7a88e00b88cf5651f2025-08-20T02:26:08ZengTaylor & Francis GroupBioTechniques0736-62051940-98182020-04-0168420020310.2144/btn-2019-0126A simple and reliable method for determination of optimum pH in coupled enzyme assaysLee Bowman0Rachael Motamed1Paul Lee2Kadijah Aleem3Astha S Berawala4Katherine L Hayden5David J Bzik6Debasish Chattopadhyay71Department of Chemistry & Physics, Birmingham-Southern College, Birmingham, AL 35254, USA1Department of Chemistry & Physics, Birmingham-Southern College, Birmingham, AL 35254, USA2Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA2Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA1Department of Chemistry & Physics, Birmingham-Southern College, Birmingham, AL 35254, USA1Department of Chemistry & Physics, Birmingham-Southern College, Birmingham, AL 35254, USA3Department of Microbiology & Immunology, Dartmouth Geisel School of Medicine, Lebanon, NH 03756, USA2Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USADetermination 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.https://www.future-science.com/doi/10.2144/btn-2019-0126coupled enzyme assayCryptosporidium parvumenzyme kineticslactate dehydrogenasepH optimizationPlasmodium falciparum
spellingShingle Lee Bowman
Rachael Motamed
Paul Lee
Kadijah Aleem
Astha S Berawala
Katherine L Hayden
David J Bzik
Debasish Chattopadhyay
A simple and reliable method for determination of optimum pH in coupled enzyme assays
BioTechniques
coupled enzyme assay
Cryptosporidium parvum
enzyme kinetics
lactate dehydrogenase
pH optimization
Plasmodium falciparum
title A simple and reliable method for determination of optimum pH in coupled enzyme assays
title_full A simple and reliable method for determination of optimum pH in coupled enzyme assays
title_fullStr A simple and reliable method for determination of optimum pH in coupled enzyme assays
title_full_unstemmed A simple and reliable method for determination of optimum pH in coupled enzyme assays
title_short A simple and reliable method for determination of optimum pH in coupled enzyme assays
title_sort simple and reliable method for determination of optimum ph in coupled enzyme assays
topic coupled enzyme assay
Cryptosporidium parvum
enzyme kinetics
lactate dehydrogenase
pH optimization
Plasmodium falciparum
url https://www.future-science.com/doi/10.2144/btn-2019-0126
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