The effects of varying the enzyme-to-substrate ratios on reaction velocity and the effect of water activity on the activation energy for the reaction could not be explained by a simple diffusional model, but required postulates that were more complex:
1. The diffusional resistance is localized in a shell adjacent to the enzyme.
2. At low water activity, the reduced hydration produces conformational changes in the enzyme, affecting its catalytic activity.
The relationship between water content and water activity is complex. An increase in aw is usually accompanied by an increase in water content, but in a non-linear fashion. This relationship between water activity and moisture content at a given temperature is called the moisture sorption isotherm. These curves are determined experimentally and constitute the fingerprint of a food system.