Cytosolic and non-cytosolic carbonic anhydrase enzymes from bovine leucocytes

In this study, carbonic anhydrase (CA) enzyme has been purified and separat ely characterized according to bound form in 4 steps as outer peripheral, c ytosolic, inner peripheral, and integral from bovine leukocyte. Affinity chromatography has also been used for purification of the enzyme i n four steps. CA has been found for each step. Measurment of enzyme activit y has been done by CO2 hydratase activity and esterase activity methods. Optimum pH and optimum temperature have been defined for each step of purif ied enzyme. The behaviors of CA with specific inhibitors, such as KSCN and NaN3 have been investigated. In each step, molecular weight and purity have been determined by gel filtration and SDS-PAGE electrophoresis. In additio n, enzyme K-M and V-max values have been determined with the method of Line weaver-Burk.