Characteristics of unbuffered gel-immobilized urease particles. I. Internal pH.

The overall rate of reaction of a gel-immobilized urease particle necessarily depends upon the hydrogen ion concentrations within the particle. When the particle is unbuffered, the internal hydrogen ion concentrations are a consequence of the local rates of reaction and the rate of egress of the products of hydrolysis. A simple apparatus has been devised which allows a fairly rapid determination of the hydrogen ion concentration in the center of a particle for any given size, enzyme concentration, substrate concentration, and external pH. The products of urea hydrolysis are self-buffering in the region of pH 8.83 and for an external pH less than the self-buffering pH, the pH within the particle is increased because of the reaction. When the external pH is greater than the self-buffering pH, the converse occurs. The pH at the center of the particle approaches the self-buffering pH with an increase in particle size and enzyme concentration. The external increase in the external substrate concentration has a limited effect, simply rendering the local rates of reaction to be of zero order. The center-line pH and therefore all internal hydrogen ion concentrations depend upon the parameter L square root pe and the external pH. Differences between the external and center-line pH values of the order of units are unexceptional. The implications of the internal pH profiles on the local and overall rates of reaction are explored.