Isolation and Purification of Arylesterase From Normal Blood Serum(Part II)

The research included isolation and purification of arylesterase from normal blood serum.The molecular weight of the partially purified enzyme was determined by gel filtration using (Sephadex G75) and SDS electrophoersis techniques. It was found that the first proteinous peak separated from ammonium sulfate saturation of serum with (25 50%) using gel filtration and SDS electrophoresis techniques had apparent molecular weight of (48123 d) and (49185d) respectively.The second proteinous peak separated from serum by gel filtration technique had apparent molecular weight (47305 d).On the other hand, the first proteinous peak separated from HDL serum had a molecular weight (49907 d).
The optimum conditions of arylesterase for the two peaks separated from HDL and ammonium sulfate precipitaion solution showed an optimum reaction incubation time at (12),(10) minutes, an optimum pH at(7.2),(7.6), an optimum temperature(30 C), (25 C) and optimum substrate concentration (4 mM), (4.5 mM) and Vmax value (243.9U/ml), (238.1 U/ml) and Km value (3.07 mM), (3.26 mM) respectively.