Metabolic isotopomer labeling systems. Part II: structural flux identifiability analysis.

Metabolic flux analysis using carbon labeling experiments (CLEs) is an important tool in metabolic engineering where the intracellular fluxes have to be computed from the measured extracellular fluxes and the partially measured distribution of 13C labeling within the intracellular metabolite pools. The relation between unknown fluxes and measurements is described by an isotopomer labeling system (ILS) (see Part I [Math. Biosci. 169 (2001) 173]). Part II deals with the structural flux identifiability of measured ILSs in the steady state. The central question is whether the measured data contains sufficient information to determine the unknown intracellular fluxes. This question has to be decided a priori, i.e. before the CLE is carried out. In structural identifiability analysis the measurements are assumed to be noise-free. A general theory of structural flux identifiability for measured ILSs is presented and several algorithms are developed to solve the identifiability problem. In the particular case of maximal measurement information, a symbolical algorithm is presented that decides the identifiability question by means of linear methods. Several upper bounds of the number of identifiable fluxes are derived, and the influence of the chosen inputs is evaluated. By introducing integer arithmetic this algorithm can even be applied to large networks. For the general case of arbitrary measurement information, identifiability is decided by a local criterion. A new algorithm based on integer arithmetic enables an a priori local identifiability analysis to be performed for networks of arbitrary size. All algorithms have been implemented and flux identifiability is investigated for the network of the central metabolic pathways of a microorganism. Moreover, several small examples are worked out to illustrate the influence of input metabolite labeling and the paradox of information loss due to network simplification.