Abstract

The topology of metabolic networks is recognisably modular with modules weakly connected apart from sharing a pool of currency metabolites. Here, we defined modules as sets of reversible reactions isolated from the rest of metabolism by irreversible reactions except for the exchange of currency metabolites. Our approach identifies topologically independent modules under specific conditions associated with different metabolic functions. As case studies, the E.coli iJO1366 and Human Recon 2.2 genome-scale metabolic models were split in 103 and 321 modules respectively, displaying significant correlation patterns in expression data. Finally, we addressed a fundamental question about the metabolic flexibility conferred by reversible reactions: Of all Directed Topologies (DTs) defined by fixing directions to all reversible reactions, how many are capable of carrying flux through all reactions?. Enumeration of the DTs for iJO1366 model was performed using an efficient depth-first search algorithm, rejecting infeasible DTs based on mass-imbalanced and loopy flux patterns. We found the direction of 79% of reversible reactions must be defined before all directions in the network can be fixed, granting a high degree of flexibility. Author summary Genome-scale metabolic reconstructions represent all biochemical reactions that an organism can accomplish. These reconstructions are complex and often difficult to study in great detail. A way to overcome this limitation is to focus on specific pathways or subsystems. We present a novel method to identify metabolic modules based on the network topology. The method relies on reaction directions and ignores currency metabolites, which artificially connect distant metabolic reactions. In this way, topologically independent modules are built, where inputs and outputs are controlled by irreversible reactions. The method is automatic and unbiased, and, the result is a set of condition specific modules with defined metabolic functions. As a proof-of-concept we generated biologically relevant modules for the E.coli and Human genome-scale metabolic reconstructions supported by transcriptomic data. Finally, we applied the novel approach to study the network flexibility conferred by reversible reactions. In the case of the E. coli model, we found that the direction of 79% of structurally reversible reactions (those not directionally constrained by surrounding irreversible reactions) must be fixed to determine all the reaction directions in the network. Therefore, reversible reactions operate practically independent of each other.