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Parallel labeling experiments and metabolic flux analysis: Past, present and future methodologies

Paper ID Volume ID Publish Year Pages File Format Full-Text
31589 44823 2013 12 PDF Available
Title
Parallel labeling experiments and metabolic flux analysis: Past, present and future methodologies
Abstract

Radioactive and stable isotopes have been applied for decades to elucidate metabolic pathways and quantify carbon flow in cellular systems using mass and isotope balancing approaches. Isotope-labeling experiments can be conducted as a single tracer experiment, or as parallel labeling experiments. In the latter case, several experiments are performed under identical conditions except for the choice of substrate labeling. In this review, we highlight robust approaches for probing metabolism and addressing metabolically related questions though parallel labeling experiments. In the first part, we provide a brief historical perspective on parallel labeling experiments, from the early metabolic studies when radioisotopes were predominant to present-day applications based on stable-isotopes. We also elaborate on important technical and theoretical advances that have facilitated the transition from radioisotopes to stable-isotopes. In the second part of the review, we focus on parallel labeling experiments for 13C-metabolic flux analysis (13C-MFA). Parallel experiments offer several advantages that include: tailoring experiments to resolve specific fluxes with high precision; reducing the length of labeling experiments by introducing multiple entry-points of isotopes; validating biochemical network models; and improving the performance of 13C-MFA in systems where the number of measurements is limited. We conclude by discussing some challenges facing the use of parallel labeling experiments for 13C-MFA and highlight the need to address issues related to biological variability, data integration, and rational tracer selection.

► We review the history of isotopic tracer experiments. ► Transition from radioisotopes to stable isotopes for flux analysis. ► Application of parallel labeling experiments for metabolism research. ► Methods for optimal selection of isotopic tracers and data integration. ► We discuss issues related to biological variability.

Keywords
ED, Entner–Doudoroff; EMP, Embden–Meyerhof–Parnas; EMU, elementary metabolite unit; EMU-BV, elementary metabolite unit basis vector; GC–MS, gas chromatography–mass spectrometry; MFA, metabolic flux analysis; MS, mass spectrometry; MS/MS, tandem mass spect
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Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 16, March 2013, Pages 21–32
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us