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Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology

(13)C-Metabolic flux analysis ((13)C-MFA) traditionally assumes that kinetic isotope effects from isotopically labeled compounds do not appreciably alter cellular growth or metabolism, despite indications that some biochemical reactions can be non-negligibly impacted. Here, populations of Escherichi...

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Autores principales: Sandberg, Troy E., Long, Christopher P., Gonzalez, Jacqueline E., Feist, Adam M., Antoniewicz, Maciek R., Palsson, Bernhard O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4786092/
https://www.ncbi.nlm.nih.gov/pubmed/26964043
http://dx.doi.org/10.1371/journal.pone.0151130
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author Sandberg, Troy E.
Long, Christopher P.
Gonzalez, Jacqueline E.
Feist, Adam M.
Antoniewicz, Maciek R.
Palsson, Bernhard O.
author_facet Sandberg, Troy E.
Long, Christopher P.
Gonzalez, Jacqueline E.
Feist, Adam M.
Antoniewicz, Maciek R.
Palsson, Bernhard O.
author_sort Sandberg, Troy E.
collection PubMed
description (13)C-Metabolic flux analysis ((13)C-MFA) traditionally assumes that kinetic isotope effects from isotopically labeled compounds do not appreciably alter cellular growth or metabolism, despite indications that some biochemical reactions can be non-negligibly impacted. Here, populations of Escherichia coli were adaptively evolved for ~1000 generations on uniformly labeled (13)C-glucose, a commonly used isotope for (13)C-MFA. Phenotypic characterization of these evolved strains revealed ~40% increases in growth rate, with no significant difference in fitness when grown on either labeled ((13)C) or unlabeled ((12)C) glucose. The evolved strains displayed decreased biomass yields, increased glucose and oxygen uptake, and increased acetate production, mimicking what is observed after adaptive evolution on unlabeled glucose. Furthermore, full genome re-sequencing revealed that the key genetic changes underlying these phenotypic alterations were essentially the same as those acquired during adaptive evolution on unlabeled glucose. Additionally, glucose competition experiments demonstrated that the wild-type exhibits no isotopic preference for unlabeled glucose, and the evolved strains have no preference for labeled glucose. Overall, the results of this study indicate that there are no significant differences between (12)C and (13)C-glucose as a carbon source for E. coli growth.
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spelling pubmed-47860922016-03-23 Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology Sandberg, Troy E. Long, Christopher P. Gonzalez, Jacqueline E. Feist, Adam M. Antoniewicz, Maciek R. Palsson, Bernhard O. PLoS One Research Article (13)C-Metabolic flux analysis ((13)C-MFA) traditionally assumes that kinetic isotope effects from isotopically labeled compounds do not appreciably alter cellular growth or metabolism, despite indications that some biochemical reactions can be non-negligibly impacted. Here, populations of Escherichia coli were adaptively evolved for ~1000 generations on uniformly labeled (13)C-glucose, a commonly used isotope for (13)C-MFA. Phenotypic characterization of these evolved strains revealed ~40% increases in growth rate, with no significant difference in fitness when grown on either labeled ((13)C) or unlabeled ((12)C) glucose. The evolved strains displayed decreased biomass yields, increased glucose and oxygen uptake, and increased acetate production, mimicking what is observed after adaptive evolution on unlabeled glucose. Furthermore, full genome re-sequencing revealed that the key genetic changes underlying these phenotypic alterations were essentially the same as those acquired during adaptive evolution on unlabeled glucose. Additionally, glucose competition experiments demonstrated that the wild-type exhibits no isotopic preference for unlabeled glucose, and the evolved strains have no preference for labeled glucose. Overall, the results of this study indicate that there are no significant differences between (12)C and (13)C-glucose as a carbon source for E. coli growth. Public Library of Science 2016-03-10 /pmc/articles/PMC4786092/ /pubmed/26964043 http://dx.doi.org/10.1371/journal.pone.0151130 Text en © 2016 Sandberg et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sandberg, Troy E.
Long, Christopher P.
Gonzalez, Jacqueline E.
Feist, Adam M.
Antoniewicz, Maciek R.
Palsson, Bernhard O.
Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title_full Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title_fullStr Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title_full_unstemmed Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title_short Evolution of E. coli on [U-(13)C]Glucose Reveals a Negligible Isotopic Influence on Metabolism and Physiology
title_sort evolution of e. coli on [u-(13)c]glucose reveals a negligible isotopic influence on metabolism and physiology
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4786092/
https://www.ncbi.nlm.nih.gov/pubmed/26964043
http://dx.doi.org/10.1371/journal.pone.0151130
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