Matrix method for determining steps most rate-limiting to metabolic fluxes in biotechnological processes.

The metabolic control theory developed by Kacser, Burns, Heinrich, and Rapoport is briefly outlined, extended, and transformed so as optimally to address some biotechnological questions. The extensions include (i) a new theorem that relates the control of metabolite concentrations by enzyme activities to flux ratios at branches in metabolic pathways; (ii) a new theorem that does the same for the control of the distribution of the flux over two branches; (iii) a method that expresses these controls into properties (the so-called elasticity coefficients) of the enzymes in the pathway; and (iv) a theorem that relates the effects of changes in metabolite concentrations on reaction rates to the effects of changes in enzyme properties on the same rates. Matrix equations relating the flux control and concentration control coefficients to the elasticity coefficients of enzymes in simple linear and branched pathways incorporating feedback are given, together with their general solutions and a numerical example. These equations allow one to develop rigorous criteria by which to decide the optimal strategy for the improvement of a microbial process. We show how this could be used in deciding which property of which enzyme should be changed in order to obtain the maximal concentration of a metabolite or the maximal metabolic flux.