The use of aqueous alkali for the titration of esterolytic activity when the esters are dissolved in alcoholic solutions, results in an error due to changes in the ionization of the buffer. This is corrected by titrating with alkali in the same solvent as the substrate.
Alcohols and other substances which change the dielectric strength of water modify the rate of hydrolysis of BAEE1 and TSAME by trypsin to an extent proportionate to their effect on the dielectric strength. The reaction rate increases with diminished dielectric strength and vice versa. At low concentrations of substance there seems to be no specific effect other than that derived of the variation in dielectric strength. At higher concentrations, the enzyme might be denatured. In addition, it is probable that specific effects of each substance might intervene. The Coulombic and thermic energies of activation were calculated for the two esters in various solvents. The plot of the logarithm of rate constant vs. reciprocal of dielectric constant yields a straight line with positive slope. This behavior is similar to that of a non-enzymatic positive ion-dipole reaction. Trypsin reacts like a positive ion. The possible influence of the dielectric strength on the regulation of the equilibria involved in the interconversion of the various forms of trypsin in solution (active, inactive, denatured) is discussed.
