Removing both Internal and Unrealistic Energy-Generating Cycles in Flux Balance Analysis

Abstract: Constraint-based stoichiometric models are ubiquitous in metabolic research, with Flux Balance Analysis (FBA) being the most widely used method to describe metabolic phenotypes of cells growing in steady-state. Of the many variants of constrain-based modelling methods published throughout the years, only few have focused on thermodynamic issues, in particular the elimination of non-physical and non-physiological cyclic fluxes. In this work, we revisit two of these methods, namely thermodynamic FBA and loopless FBA, and analyze the strengths and weaknesses of each one. Finally, we suggest a compromise denoted semi-thermodynamic FBA (st-FBA) which imposes stronger thermodynamic constrains on the flux polytope compared to loopless FBA, without requiring a large set of thermodynamic parameters as in the case of thermodynamic FBA. We show that st-FBA is a useful and simple way to eliminate thermodynamically infeasible cycles that generate ATP.