The optimum temperature of salicin hydrolysis by enzyme action is independent of the concentrations of substrate and enzyme
The object of the present investigation is to ascertain the influence, if any, on the optimum temperature—temperature of greatest activity—of an enzyme, of the concentration, on the one hand, of the substrate, and, on the other, of the enzyme. The investigation, involving two variables, presents three cases for consideration, according as the concentration of the substrate and the concentration of the enzyme are varied separately or together. An account is given of the results obtained with the enzyme or enzymic function, present in sweet-almond emulsin, which hydrolyses the glucoside salicin with the production of equimolecular quantities of glucose and saligenin. A commercial specimen of Merck’s emulsin was used, while the purity of the salicin employed was ascertained by determining its melting point (200-5°) and its optical activity ([ α ] 21 D = -62·7°). The successive stages in the inquiry may be briefly summarised as follows: (1) a preliminary determination of the activity of the specimen under certain chosen conditions as regards the concentration of the substrate, the temperature, and the duration of the experiment; (2) a preliminary determination of the optimum temperature with the quantity of enzyme found capable of producing 50 per cent, hydrolysis of the substrate under the above conditions; (3) a determination of the activity curves of the enzyme at the temperature thus found, in an action of the same duration for five concentrations of the substrate M/5, M/10, M/15, M/30, and M/50; (4) a determination of the optimum temperature of the enzyme for each of the five concentrations of the substrate in presence of a constant enzyme concentration ; (5) a determination of the optimum temperature of the enzyme for each of the five concentrations of the substrate with quantities of enzyme indicated by the activity curves as capable of producing 70 per cent, hydrolysis of the substrate in the given time: (6) a determination of the optimum temperature of the enzyme for a constant concentration of the substrate in presence of different enzyme concentrations.