Karbonik anhidraz enziminin afinite kromatografisiyle saflaştirilmasi ve biyoteknolojik uygulamalari

In this study, the carbonic anhydrase (CA) enzyme has been purified with a new affinity support developed from sheep liver. Within the scope of the study; the interaction between the sulfonamide, one of the inhibitors of CA enzyme, and threonine amino acid, has been mimicked and cryogel column has been synthesized in the presence of the sulfonamides and MA-threonine pre-organization with cross-proper linkers. Characterization of the synthesized cryogel column has been performed by swelling test, BET, FT-IR and SEM analysis. In the next step of the study, the purification of CA enzyme has been realized by Fast Protein Liquid Chromatography (FPLC) from crude extract obtained from sheep liver. The protein content of the purified enzyme has been determined by Bradford method, UV spectrometric measurements have been showed the optimum conditions for esterase and hydratase activity, Circular Dichroism (CD) Spectroscopy has been used to evaluate changes in secondary and tertiary structure and molecular weight has been determined by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Keywords: Carbonic anhydrase, cryogel column, sulphonamide, threonine, mimic interaction