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Related Concept Videos

Carbon-13 (¹³C) NMR: Overview01:10

Carbon-13 (¹³C) NMR: Overview

Carbon-13 is a naturally occurring NMR-active isotope of carbon with a low natural abundance of 1.1%. In contrast, carbon-12 is the most abundant isotope of carbon with zero nuclear spin. Therefore, it is NMR inactive. The gyromagnetic ratio of carbon-13 is smaller than that of protons. As a result, carbon-13 resonance is about 6000 times weaker than proton resonance. For a given magnetic field strength, the resonance frequency of carbon-13 is about one-fourth of the resonance frequency for...
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Protons in identical electronic environments within a molecule are chemically equivalent and have the same chemical shift. The replacement test is a useful tool to identify chemical equivalence and predict NMR spectra. A substituent replaces each of the protons being examined and the resulting molecules are compared. If the same molecule is obtained, the protons are equivalent or homotopic. Replacement of any hydrogens in ethane by chlorine yields chloroethane because all six protons are...
¹³C NMR: ¹H–¹³C Decoupling01:04

¹³C NMR: ¹H–¹³C Decoupling

The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...

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Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling
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Stationary versus non-stationary (13)C-MFA: a comparison using a consistent dataset.

Stephan Noack1, Katharina Nöh, Matthias Moch

  • 1Institut für Biotechnologie, Forschungszentrum Jülich, Leo-Brandt-Straße, Jülich, NRW, Germany. k.noeh@fz-juelich.de

Journal of Biotechnology
|July 20, 2010
PubMed
Summary

This study compares two methods for analyzing cellular metabolism: stationary and non-stationary ¹³C-metabolic flux analysis (MFA). Results show that fluxome data significantly depends on the chosen method and experimental setup.

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Area of Science:

  • Metabolic Engineering
  • Systems Biology
  • Biotechnology

Background:

  • ¹³C-metabolic flux analysis (MFA) is crucial for understanding cellular metabolism.
  • Isotopically non-stationary MFA offers faster fluxome analysis compared to traditional stationary methods.
  • Direct comparison of both MFA approaches using a single experimental dataset was lacking.

Purpose of the Study:

  • To directly compare stationary and non-stationary ¹³C-MFA using data from a single experiment.
  • To evaluate the impact of measurement and modeling on fluxome data.
  • To identify limitations and potential pitfalls of both MFA methods.

Main Methods:

  • A ¹³C-labeling experiment was conducted on a *Corynebacterium glutamicum* lysine producer during fed-batch cultivation.
  • Substrate glucose was switched from unlabeled to labeled, with rapid sampling and metabolite quenching.
  • Intracellular metabolite labeling enrichments were monitored over time using LC-MS/MS and analyzed with both stationary and non-stationary ¹³C-MFA.

Main Results:

  • Quantitative fluxome data derived from both methods were found to be significantly dependent on the measurement strategy and modeling approach.
  • Relative flux distributions and quantitative fluxome data varied based on the MFA method employed.
  • The study highlighted experimental and computational limitations inherent to both stationary and non-stationary ¹³C-MFA.

Conclusions:

  • The choice of ¹³C-MFA method and experimental design critically influences the resulting metabolic flux profiles.
  • Understanding the limitations of each approach is essential for accurate fluxome characterization.
  • This work provides a basis for optimizing experimental strategies and data integration in metabolic flux analysis.