The Influence of the Substrate Concentration on the Rate of Hydrolysis of Proteins by Pepsin

1. It is pointed out that the apparent exceptions to the law of mass action found in enzyme reactions may be found in catalytic reactions in strictly homogeneous solutions. 2. These deviations in the rate of reaction from the law of mass action may be explained by the hypothesis that the active mass of the reacting substances is not directly proportional to the total concentration of substance taken. 3. In support of this suggestion it is shown that for any given concentration of pepsin the relative rate of digestion of concentrated and of dilute protein solutions is always the same. If the rate of digestion depended on the saturation of the surface of the enzyme by substrate the relative rate of digestion of concentrated protein solutions should increase more rapidly with the concentration of enzyme than that of dilute solutions. This was found not to be true, even when the enzyme could not be considered saturated in the dilute protein solutions. 4. The rate of digestion and the conductivity of egg albumin solutions of different concentration were found to be approximately proportional at the same pH. This agrees with the hypothesis first expressed by Pauli that the ionized protein is largely or entirely the form which is attacked by the enzyme. 5. The rate of digestion is diminished by a very large increase in the viscosity of the protein solution. This effect is probably a mechanical one due to the retardation of the diffusion of the enzyme.