Isotopically nonstationary metabolic flux analysis (INST-MFA): putting theory into practice

Cheah, Yi Ern; Young, Jamey D.

doi:10.1016/j.copbio.2018.02.013

Isotopically nonstationary metabolic flux analysis (INST-MFA): putting theory into practice

Journal Article · Mon Mar 05 23:00:00 EST 2018 · Current Opinion in Biotechnology

DOI:https://doi.org/10.1016/j.copbio.2018.02.013· OSTI ID:1611014

Cheah, Yi Ern ^[1]; Young, Jamey D. ^[2]

Vanderbilt Univ., Nashville, TN (United States); DOE/OSTI
Vanderbilt Univ., Nashville, TN (United States)

Typically, ¹³C flux analysis relies on assumptions of both metabolic and isotopic steady state. If metabolism is steady but isotope labeling is not allowed to fully equilibrate, isotopically nonstationary metabolic flux analysis (INST-MFA) can be used to estimate fluxes. This requires solution of differential equations that describe the time-dependent labeling of network metabolites, while iteratively adjusting the flux and pool size parameters to match the transient labeling measurements. INST-MFA holds a number of unique advantages over approaches that rely solely upon steady-state isotope enrichments. First, INST-MFA can be applied to estimate fluxes in autotrophic systems, which consume only single-carbon substrates. Second, INST-MFA is ideally suited to systems that label slowly due to the presence of large intermediate pools or pathway bottlenecks. Finally, INST-MFA provides increased measurement sensitivity to estimate reversible exchange fluxes and metabolite pool sizes, which represents a potential framework for integrating metabolomic analysis with ¹³C flux analysis. This review highlights the unique capabilities of INST-MFA, describes newly available software tools that automate INST-MFA calculations, presents several practical examples of recent INST-MFA applications, and discusses the technical challenges that lie ahead.

View Accepted Manuscript (DOE)

Research Organization:: Vanderbilt Univ., Nashville, TN (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: SC0008118

OSTI ID:: 1611014

Alternate ID(s):: OSTI ID: 1778466

Journal Information:: Current Opinion in Biotechnology, Journal Name: Current Opinion in Biotechnology Vol. 54; ISSN 0958-1669

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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