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pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions
Under carbon source transitions, the intracellular pH of Saccharomyces cerevisiae is subject to change. Dynamics in pH modulate the activity of the glycolytic enzymes, resulting in a change in glycolytic flux and ultimately cell growth. To understand how pH affects the global behavior of glycolysis...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9790636/ https://www.ncbi.nlm.nih.gov/pubmed/35429225 http://dx.doi.org/10.1111/febs.16459 |
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author | Luzia, Laura Lao‐Martil, David Savakis, Philipp van Heerden, Johan van Riel, Natal Teusink, Bas |
author_facet | Luzia, Laura Lao‐Martil, David Savakis, Philipp van Heerden, Johan van Riel, Natal Teusink, Bas |
author_sort | Luzia, Laura |
collection | PubMed |
description | Under carbon source transitions, the intracellular pH of Saccharomyces cerevisiae is subject to change. Dynamics in pH modulate the activity of the glycolytic enzymes, resulting in a change in glycolytic flux and ultimately cell growth. To understand how pH affects the global behavior of glycolysis and ethanol fermentation, we measured the activity of the glycolytic and fermentative enzymes in S. cerevisiae under in vivo‐like conditions at different pH. We demonstrate that glycolytic enzymes exhibit differential pH dependencies, and optima, in the pH range observed during carbon source transitions. The forward reaction of GAPDH shows the highest decrease in activity, 83%, during a simulated feast/famine regime upon glucose removal (cytosolic pH drop from 7.1 to 6.4). We complement our biochemical characterization of the glycolytic enzymes by fitting the V(max) to the progression curves of product formation or decay over time. The fitting analysis shows that the observed changes in enzyme activities require changes in V(max) , but changes in K(m) cannot be excluded. Our study highlights the relevance of pH as a key player in metabolic regulation and provides a large set of quantitative data that can be explored to improve our understanding of metabolism in dynamic environments. |
format | Online Article Text |
id | pubmed-9790636 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-97906362022-12-28 pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions Luzia, Laura Lao‐Martil, David Savakis, Philipp van Heerden, Johan van Riel, Natal Teusink, Bas FEBS J Original Articles Under carbon source transitions, the intracellular pH of Saccharomyces cerevisiae is subject to change. Dynamics in pH modulate the activity of the glycolytic enzymes, resulting in a change in glycolytic flux and ultimately cell growth. To understand how pH affects the global behavior of glycolysis and ethanol fermentation, we measured the activity of the glycolytic and fermentative enzymes in S. cerevisiae under in vivo‐like conditions at different pH. We demonstrate that glycolytic enzymes exhibit differential pH dependencies, and optima, in the pH range observed during carbon source transitions. The forward reaction of GAPDH shows the highest decrease in activity, 83%, during a simulated feast/famine regime upon glucose removal (cytosolic pH drop from 7.1 to 6.4). We complement our biochemical characterization of the glycolytic enzymes by fitting the V(max) to the progression curves of product formation or decay over time. The fitting analysis shows that the observed changes in enzyme activities require changes in V(max) , but changes in K(m) cannot be excluded. Our study highlights the relevance of pH as a key player in metabolic regulation and provides a large set of quantitative data that can be explored to improve our understanding of metabolism in dynamic environments. John Wiley and Sons Inc. 2022-05-11 2022-10 /pmc/articles/PMC9790636/ /pubmed/35429225 http://dx.doi.org/10.1111/febs.16459 Text en © 2022 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Luzia, Laura Lao‐Martil, David Savakis, Philipp van Heerden, Johan van Riel, Natal Teusink, Bas pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title | pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title_full | pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title_fullStr | pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title_full_unstemmed | pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title_short | pH dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
title_sort | ph dependencies of glycolytic enzymes of yeast under in vivo‐like assay conditions |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9790636/ https://www.ncbi.nlm.nih.gov/pubmed/35429225 http://dx.doi.org/10.1111/febs.16459 |
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