Abstract
Stable isotope labelling experiments provide many opportunities for probing metabolic pathways. One goal of such experiments is to define the metabolic phenotype of an organism in terms of the fluxes supported by the entire metabolic network. Metabolic flux analysis (MFA) is used routinely in prokaryotes, but its application to plants is more challenging. Here we examine the status of MFA in plants, highlighting difficulties that hinder wider exploitation of the technique. We conclude that simulation of network fluxes using constraints-based modelling offers a more versatile approach for exploring the capabilities of a network, and that MFA might be best used to probe the interesting features that simulation reveals.
Highlights
Stable isotope labelling experiments provide many opportunities for probing metabolic pathways. One goal of such experiments is to define the metabolic phenotype of an organism in terms of the fluxes supported by the entire metabolic network
If an experimental system in a metabolic steady state can be labelled to an isotopic steady state, so-called steady-state metabolic analysis can be used to construct a flux map for the principal pathways of central metabolism (Box1)
Production in a heterotrophic Arabidopsis thaliana cell culture, the interpretation was confounded by the high rates of the exchange reactions catalysed by flavin-containing enzymes (Smith et al, 2019)
Summary
Analysing labelling time-courses in a metabolic steady state using the technique known as non-stationary MFA (INST-MFA) (Wiechert and Nöh, 2013) is a potential solution to this problem, but so far there have been just two applications to leaves (Ma et al, 2014; Xu et al., 2021) These studies demonstrated that INST-MFA can provide a system-wide analysis of the metabolic fluxes in photoautotrophic metabolic networks, together with quantitative information on the pool sizes of metabolic intermediates.
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