Determination of carbonic anhydrase (E.C. 4.2.1.1.) activity by means of the pH-stat-technique.

An improved pH-stat-technique for the determination of carbonic anhydrase is presented. As compared with common techniques the main advantages are: 1. The hydration reaction of CO2 catalyzed by carbonic anhydrase can be determined at a constant and selected pH-value, 2. the substrate concentration can be kept constant, 3. the reaction volume amounts to only 2 ml, and 4. the reaction can be measured continously for longer periods of time. The applicability of the method is demonstrated by several well known kinetic data. Instructions for performing the experiments and for the calculation of the results are presented in the Materials and Methods section of the paper.