| Application Notes | Experiment Notes: Carbonic anhydrase from bovine erythrocytes. The enzyme catalyzes the following reaction Carbonic anhydrase CO2+H2O H2CO3 Molecular weight of protein is about 30 kDa. Isoelectric point of protein 5.65. Molar extinction at 280 nm is 5700 Mol-1cm-1. Activity is about 2500 W-A U/mg protein. Assay: Reaction mixture contains 2 ml 0.025 M veronal pH 8.2, bromthymol blue 1mg/ 100 ml. Enzyme solution 1 ml is added to 2 ml of CO2 saturated buffer solution and time is registered until color of indicator is changed from blue to yellow. Activity is calculated from the following equation U=10 (Tb/Tc-1)/mg of protein Where Tb- time of noncatalyzed reaction Tc- time of enzyme catalyzed reaction. |
|---|