Purification, characterization and dertermination of kinetic features of carbonic anhydrase from turbot (Psetta Maxima) muscle tissue

In this study, carbonic anhydrase purification from turbot (Psetta maxima) muscle tissue, together with analysis of the kinetic behavior and some enzyme properties, is described. The purification steps comprised hemolysate preparation, Sepharose-4B-L tyrosine-sulfanilamide affinity gel chromatography, and dialyzing. The yield was 69.05%, and the enzyme was found to have a specific activity of 755.2 EU/mg protein. The overall purification was about 50.65-fold. A temperature of +4 °C was maintained during the purification process. The molecular mass of the subunit was determined to be 29.7 kDa by SDS polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme had an optimal pH of 8.0, a stable pH of 8.0, and an optimal temperature of 30 °C. Km and Vmax for p-nitrophenylacetate as a substrate were also determined.