Steady-state ¹³C fluxomics using OpenFLUX

Metabolic flux estimation using (13)C isotopic tracers ((13)C-MFA) provides a greater resolution of intracellular fluxes than using only cell growth and consumption/production rates. However, (13)C-MFA is computationally more demanding. A nonlinear least-square optimization process is employed to constrain metabolic fluxes using atom balance models and experimentally measured (13)C labelling pattern of intracellular or proteinogenic metabolites. OpenFLUX was therefore developed for the purpose of streamlining the computational workflow. Here, we describe in detail the computational procedure for performing (13)C-MFA using OpenFLUX. We also provide some helpful information on model reconstruction and GC-MS data treatment.