Real-time detection of central carbon metabolism in living<i>Escherichia coli</i>and its response to perturbations

Meier, Sebastián; Jensen, Pernille Rose; Duus, Jens Øllgaard

doi:10.1016/j.febslet.2011.08.049

Cited by 65 publications

(98 citation statements)

References 29 publications

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“…Spin-Lattice Relaxation Times (T 1 ) and the T 1 (1)/T 1 (2) Ratios for Carbon Signals in 13 C 6 -Glucose-d 7 (1) and a) Obtained from inversion recovery method using a 11.4T NMR spectroscopy. ized 1 using Escherichia coli and yeast reported by Meier and coworkers, [41][42][43][44] signals of several metabolites containing phosphorylated β-fructofuranoses generated through glycolysis, pentose phosphate pathway, and alcohol fermentation are detected. To analyze these metabolic differences in living microorganisms observed directly by feeding the hyperpolarized glucose can lead to understand the phenotypic and genotypic differences.…”

Section: Resultsmentioning

confidence: 99%

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

Kumagai

Akakabe

Tsuda

et al. 2014

Biological & Pharmaceutical Bulletin

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Hyperpolarization of stable isotope-labeled substrates and subsequent NMR measurement of the metabolic reactions allow for direct tracking of cellular reactions in vitro and in vivo. Here, we report the hyperpolarization of 13 C 6 -glucose-d 7 and evaluate its use as probes to observe glucose flux in cells. We measured the lifetime of the polarized signal governed by the spin-lattice relaxation time T 1 .13 C 6 -Glucose-d 7 exhibited a T 1 that was over ten times as long as that of 13 C 6 -glucose, and metabolic NMR studies of hyperpolarized 13 C 6 -glucose-d 7 using tumor cell lysate led to observation of the resonances due to phosphorylated fluctofuranoses generated through aerobic glycolysis.

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Section: Resultsmentioning

confidence: 99%

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

Kumagai

Akakabe

Tsuda

et al. 2014

Biological & Pharmaceutical Bulletin

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“…Since T 1 decay of 13 C polarization limits the duration of data acquisition, incorporation of 2 H prolongs the T 1 of 13 C in some molecules, enabling a longer period of observation [2]. Metabolism of hyperpolarized [U- 2 H 7 , U- 13 C 6 ]glucose to hyperpolarized [U- 13 C 3 ]lactate has been described in Escherichia coli [3], yeast [4], breast cancer cells [5] and lymphoma [6]. These findings are important because imaging of hyperpolarized lactate derived from glucose could provide a direct measure of glycolysis and thereby offer a fundamentally new method for investigating carbohydrate metabolism in vivo [6].…”

Section: Introductionmentioning

confidence: 99%

The rate of lactate production from glucose in hearts is not altered by per-deuteration of glucose

Funk

Anderson

Wen

et al. 2017

Journal of Magnetic Resonance

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This study was designed to determine whether perdeuterated glucose experiences a kinetic isotope effect (KIE) as glucose passes through glycolysis and is further oxidized in the tricarboxylic acid (TCA) cycle. Metabolism of deuterated glucose was investigated in two groups of perfused rat hearts. The control group was supplied with a 1:1 mixture of [U-13C6]glucose and [1,6-13C2]glucose, while the experimental group received [U-13C6,U-2H7]glucose and [1,6-13C2]glucose. Tissue extracts were analyzed by 1H, 2H and proton-decoupled 13C NMR spectroscopy. Extensive 2H-13C scalar coupling plus chemical shift isotope effects were observed in the proton-decoupled 13C NMR spectra of lactate, alanine and glutamate. A small but measureable (~8%) difference in the rate of conversion of [U-13C6]glucose vs. [1,6-13C2]glucose to lactate, likely reflecting rates of C–C bond breakage in the aldolase reaction, but conversion of [U-13C6]glucose versus [U-13C6,U-2H7]glucose to lactate did not differ. This shows that the presence of deuterium in glucose does not alter glycolytic flux. However, there were two distinct effects of deuteration on metabolism of glucose to alanine and oxidation of glucose in the TCA. First, alanine undergoes extensive exchange of methyl deuterons with solvent protons in the alanine amino transferase reaction. Second, there is a substantial kinetic isotope effect in metabolism of [U-13C6,U-2H7]glucose to alanine and glutamate. In the presence of [U-13C6,U-2H7]glucose, alanine and lactate are not in rapid exchange with the same pool of pyruvate. These studies indicate that the appearance of hyperpolarized 13C-lactate from hyperpolarized [U-13C6,U-2H7]glucose is not substantially influenced by a deuterium kinetic isotope effect.

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“…Bacterial metabolism has been previously investigated using HP 13 C glucose in E. coli 36 . In that study, the common fermentative byproduct, ethanol, was observed.…”

Section: Discussionmentioning

confidence: 99%

Detection of Bacteria-Specific Metabolism Using Hyperpolarized [2-¹³C]Pyruvate

et al. 2018

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The differentiation of bacterial infection from other causes of inflammation is difficult in clinical practice and is critical where patient outcomes rely heavily on early interventions. In addition to physical exam and laboratory markers, several imaging modalities are frequently employed, but these techniques generally target the host immune response, rather than the living microorganisms themselves. Here, we describe a method to detect bacteria-specific metabolism using hyperpolarized (HP) 13C magnetic resonance spectroscopy. This technology allows visualization of the real-time conversion of enriched 13C substrates to their metabolic products, identified by their distinct chemical shifts. We have identified the rapid metabolism of HP [2-13C]pyruvate to [1-13C]acetate as a metabolic signature of common bacterial pathogens. We demonstrate this conversion in representative Gram negative and Gram positive bacteria, namely Escherichia coli and Staphylococcus aureus, and its absence in key mammalian cell-types. Furthermore this conversion was successfully modulated in three mutant strains, corresponding to deletions of relevant enzymes.

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Real-time detection of central carbon metabolism in livingEscherichia coliand its response to perturbations

Cited by 65 publications

References 29 publications

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

The rate of lactate production from glucose in hearts is not altered by per-deuteration of glucose

Detection of Bacteria-Specific Metabolism Using Hyperpolarized [2-¹³C]Pyruvate

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Real-time detection of central carbon metabolism in livingEscherichia coliand its response to perturbations

Cited by 65 publications

References 29 publications

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

Observation of Glycolytic Metabolites in Tumor Cell Lysate by Using Hyperpolarization of Deuterated Glucose

The rate of lactate production from glucose in hearts is not altered by per-deuteration of glucose

Detection of Bacteria-Specific Metabolism Using Hyperpolarized [2-13C]Pyruvate

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Detection of Bacteria-Specific Metabolism Using Hyperpolarized [2-¹³C]Pyruvate