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

13C-Metabolic flux analysis (13C-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 co...

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Bibliographic Details
Published in:PloS one 2016-03, Vol.11 (3), p.e0151130-e0151130
Main Authors: Sandberg, Troy E, Long, Christopher P, Gonzalez, Jacqueline E, Feist, Adam M, Antoniewicz, Maciek R, Palsson, Bernhard O
Format: Article
Language:English
Subjects:
Acetic acid
Adaptation, Biological - genetics
Analysis
Bioengineering
Biology
Biology and Life Sciences
Carbon
Carbon Radioisotopes - toxicity
Carbon sources
Chromatography
Cloning
DNA, Bacterial - chemistry
E coli
Engineering
Escherichia coli
Escherichia coli - drug effects
Escherichia coli - metabolism
Escherichia coli - physiology
Evolution
Evolution & development
Evolution, Molecular
Evolutionary adaptation
Experiments
Fitness
Gene expression
Gene sequencing
Genome, Bacterial
Genomes
Glucose
Glucose - metabolism
Growth rate
Labeling
Laboratories
Mass spectrometry
Metabolic flux
Metabolic Flux Analysis
Metabolism
Mimicry
Mutation
Oxygen
Oxygen uptake
Physical Sciences
Physiological aspects
Reproductive fitness
Research and Analysis Methods
Scientific imaging
Sequence Analysis, DNA
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Summary:13C-Metabolic flux analysis (13C-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 13C-glucose, a commonly used isotope for 13C-MFA. Phenotypic characterization of these evolved strains revealed ~40% increases in growth rate, with no significant difference in fitness when grown on either labeled (13C) or unlabeled (12C) 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 12C and 13C-glucose as a carbon source for E. coli growth.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0151130