@Article{, title={Human Carbonic Anhydrase: Purification and Characterization Study in Thalassemia Major Patients Compared to Healthy Subjects}, author={Salwa Saleh Hussein, Israa Ghassan Zainal}, journal={Medical Journal of Babylon مجلة بابل الطبية}, volume={15}, number={4}, pages={349-356}, year={2018}, abstract={Background: Carbonic anhydrase (CA) catalyzes the reversible reaction of converting carbon dioxide to bicarbonate. Objective: This studywas aimed to isolate and purify human erythrocytes CA and study its physicochemical properties of the enzyme reaction for ß‑thalassemia majorpatients. Materials and Methods: The blood samples included 61 samples of blood (31 males and 30 females) from ß‑thalassemia patientsvisited Azadi Hospital/Kirkuk city. Healthy individuals as control group included 40 participants. The separated fractions were obtained usingfour steps: extraction by ethanol and chloroform, ammonium sulfate precipitation, dialysis, and gel filtration chromatography; finally, the CAwas analyzed by polyacrylamide gel electrophoresis. Results: The CA activity showed significant (P ≤ 0.05) decrease, total protein showednonsignificant (P ≥ 0.05) increase, and specific activity significantly (P ≤ 0.05) increased in patients group compared to healthy individuals.CA was partially purified with a factor of 22.5 and 18 by extraction with ethanol and chloroform and 1.5,1.4 for Fraction I and 1,2 for FractionII using gel filtration chromatography. The optimum conditions for the CA reaction in patients group were enzyme concentration (6 μl),substrate concentration (6 Mm), pH = 7.4, and temperature 37°C. The electrophoresis study indicated that the bands of CA in patients groupshowed bands with less intensity than the bands in healthy individuals. Conclusion: The best method to purify CA from human erythrocyteswith high recovery and fold of purification was ethanol–chloroform extraction.

} }