Cytosolic and non-Cytosolic Carbonic Anhydrase Enzymes form Bovine Leucocytes

摘要

In this study, carbonic anhydrase (CA) enzyme has been purified and separately characterized according to bound from in 4 steps as outer peripheral, cytosolic, inner peripheral, and integral from bovine leukocye. Affinity chromatography has also been used for purification of the enzyme in four steps. CA has been found for each step. Measurement of enzyme activity has been done by CO_2 hydratase activity and esterase activity methods. Optimum opH and optimum temperature have been defined for each step of purified enzyme. The behaviors of CA with specific inhibitors, such as KSCN and NaN_3 have been investigated. In each step, molecular weight and purity have been determined byh gel filtration and SDS-PAGE electrophoresis. In addition, enzyme K_M and V_max values have been determined with the method of Lineweaver-Burk.