Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics

Wei Xiong; Huaiguang Jiang; PinChing Maness

doi:10.1007/978-1-0716-0195-2_14

Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics

Wei Xiong, Huaiguang Jiang, PinChing Maness

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

4 Scopus Citations

Abstract

Metabolic flux analysis represents an essential perspective to understand cellular physiology and offers quantitative information to guide pathway engineering. A valuable approach for experimental elucidation of metabolic flux is dynamic flux analysis, which estimates the relative or absolute flow rates through a series of metabolic intermediates in a given pathway. It is based on kinetic isotope labeling experiments, liquid chromatography-mass spectrometry (LC-MS), and computational analysis that relate kinetic isotope trajectories of metabolites to pathway activity. Herein, we illustrate the mathematic principles underlying the dynamic flux analysis and mainly focus on describing the experimental procedures for data generation. This protocol is exemplified using cyanobacterial metabolism as an example, for which reliable labeling data for central carbon metabolites can be acquired quantitatively. This protocol is applicable to other microbial systems as well and can be readily adapted to address different metabolic processes.

Original language	American English
Title of host publication	Metabolic Pathway Engineering
Subtitle of host publication	Methods in Molecular Biology, Volume 2096
Editors	M. E. Himmel, Y. J. Bomble
Publisher	Humana Press Inc.
Pages	179-196
Number of pages	18
DOIs	https://doi.org/10.1007/978-1-0716-0195-2_14 https://doi.org/10.1007/978-1-0716-0195-2_14
State	Published - 2020

Publication series

Name	Methods in Molecular Biology
Volume	2096
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.

NREL Publication Number

NREL/CH-2700-77590

Keywords

Cell harvesting
Dynamic flux analysis
Experimental metabolomics
Isotope tracer
LC-MS
Metabolic flux
Quenching

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Xiong, W., Jiang, H., & Maness, P. (2020). Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics. In M. E. Himmel, & Y. J. Bomble (Eds.), Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096 (pp. 179-196). (Methods in Molecular Biology; Vol. 2096). Humana Press Inc.. https://doi.org/10.1007/978-1-0716-0195-2_14, https://doi.org/10.1007/978-1-0716-0195-2_14

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Xiong, W, Jiang, H & Maness, P 2020, Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics. in ME Himmel & YJ Bomble (eds), Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096. Methods in Molecular Biology, vol. 2096, Humana Press Inc., pp. 179-196. https://doi.org/10.1007/978-1-0716-0195-2_14, https://doi.org/10.1007/978-1-0716-0195-2_14

Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics. / Xiong, Wei; Jiang, Huaiguang; Maness, PinChing.
Metabolic Pathway Engineering: Methods in Molecular Biology, Volume 2096. ed. / M. E. Himmel; Y. J. Bomble. Humana Press Inc., 2020. p. 179-196 (Methods in Molecular Biology; Vol. 2096).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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Chapter 14: Dynamic Flux Analysis: An Experimental Approach of Fluxomics

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