The dependence of the specific activity of urease upon the apparent absolute enzyme concentration

Abstract

If the activity of an enzyme preparation is determined under conditions in which a further increase in substrate concentration is without demonstrable effect, all other factors being held constant, it is ordinarily assumed that the specific activity of the enzyme, expressed in terms of arbitrary units per unit weight of the enzyme, is independent of the absolute enzyme concentration (1, 2). However, with urease solutions stabilized with hydrogen sulfide or cysteine (1) we have observed that the specific activity of a given urease preparation, when determined under the above conditions, increases with decreasing apparent enzyme concentration over a wide range of concentrations and that this increase in specific activity proceeds with a measurable velocity at temperatures above 15°. This phenomenon was observed with crude urease preparations, such as jack bean meal, and with two, three, and seven times recrystallized urease. Since little or no difference was observed in the behavior of three and seven times recrystallized urease, the data presented in this paper are limited to those obtained with thrice recrystallized preparations. Urease activity was determined by a modification of the procedure described by Van Slyke and Cullen (3). The precision of the modified procedure was ±2 to 3 per cent.