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A novel approach to metabolic network analysis using a Nash Equilibrium formulation is proposed. Enzymes are considered to be players in a multi-player game in which each player attempts to minimize the dimensionless Gibbs free energy associated with the biochemical reaction(s) it catalyzes subject to elemental mass balances. Mathematical formulation of the metabolic network as a set of nonlinear programming (NLP) sub-problems and appropriate solution methodologies are described. A small example representing part of the production cycle for acetyl-CoA is used to demonstrate the efficacy of the proposed Nash Equilibrium framework and show that it represents a paradigm shift in metabolic network analysis. A Nash Equilibrium Approach to Metabolic Network Analysis Angelo Lucia1(B) and Peter A. DiMaggio 1 Department of Chemical Engineering, University of Rhode Island, Kingston, RI 02881, USA [email protected] 2 Department of Chemical Engineering, Imperial College London, London SW7 2AZ, UK Abstract. A novel approach to metabolic network analysis using a Nash Equilibrium formulation is proposed. Enzymes are considered to be playAQ1 A novel approach to metabolic network analysis using a Nash Equilibrium formulation is proposed. Enzymes are considered to be playAQ1 ers in a multi-player game in which each player attempts to minimize the dimensionless Gibbs free energy associated with the biochemical reaction(s) it catalyzes subject to elemental mass balances. Mathematical formulation of the metabolic network as a set of nonlinear programming (NLP) sub-problems and appropriate solution methodologies are described. A small example representing part of the production cycle for acetyl-CoA is used to demonstrate the efficacy of the proposed Nash Equilibrium framework and show that it represents a paradigm shift in metabolic network analysis.