Carbonic anhydrase from bovine bone

In this research, carbonic anhydrase enzyme, which was taken from the bones of an animal, was purified and characterized for the first time. For this, the bones of a young cow were used. The purification treatment was complet ed in three steps. Three different isoenzymes, such as peripheral, cystolic , and integral from the bone-cell cytozolic isoenzyme were purified and cha racterized. In purification of the three isoenzymes, the technique of affin ity chromatography, which utilized Sepharose-4B-L-Tyrosine-Sulphanylamide, was used. In measuring the activities of enzymes, two different methods wer e applied. These are the esterase methods that utilize hydratase and p-nitr ophenylacetate as substrate. The measurement of proteins was done with the methods of Bradford and Coomassie Brillant Blue. The optimum pH and tempera ture of each enzyme were measured and molecular weights were measured by ge l-filtration. Its purity was examined by SDS-PAGE (3-10% alternating) elect rophoresis and the inferior unit was defined. The inhibition effects of som e chemicals were tested for each of the three isoenzymes.