  Enzyme-mediated reactions may proceed through multiple intermediate
conformational states before creating a final product molecule, and one often
wishes to identify such intermediate structures from observations of the
product creation. In this paper, we address this problem by solving the
chemical master equations for various enzymatic reactions. We devise a
perturbation theory analogous to that used in quantum mechanics that allows us
to determine the first (<n>) and the second (variance) cumulants of the
distribution of created product molecules as a function of the substrate
concentration and the kinetic rates of the intermediate processes. The mean
product flux V=d<n>/dt (or "dose-response" curve) and the Fano factor
F=variance/<n> are both realistically measurable quantities, and while the mean
flux can often appear the same for different reaction types, the Fano factor
can be quite different. This suggests both qualitative and quantitative ways to
discriminate between different reaction schemes, and we explore this
possibility in the context of four sample multistep enzymatic reactions. We
argue that measuring both the mean flux and the Fano factor can not only
discriminate between reaction types, but can also provide some detailed
information about the internal, unobserved kinetic rates, and this can be done
without measuring single-molecule transition events.
