Akademik Veri Yönetim Sistemi
Bioscience Research, cilt.16, sa.4, ss.3834-3845, 2019 (ESCI)
In our work, hPON1 enzyme was purified from human serum using the Three Phase Partitioning (TPP) and it was immobilized on functionalized ferric magnetic chitosan nanoparticles (MC-NPs) by the covalent binding method. The optimized amount for parameters of enzyme units (EU)/mg MC-NPs, reaction time, pH and temperature were determined to be 230(EU)/mg, 30 min, 8.0, and 37oC respectively. The amount of immobilization yield according to the enzyme activity was obtained to be 88% and also the amount of immobilized enzyme on MC-NPs was 0.23 EU/mg. Stability studies showed significant increase in immobilized enzyme stability at 4, 25 and 37°C. The stability of Immobilized enzyme showed a 6.4-fold increase in comparison to free enzyme at 37°C. The results demonstrated that the pH stability of the immobilized enzyme significantly increased in comparison with free enzyme. The immobilized enzyme was usable and recoverable for ten cycles. The results depicted that 85% of enzyme activity was retained after fifth cycle. SEM, XRD and FTIR test showed the covalent binding of enzyme to magnetic nanoparticles’ surface. In the last stage of the study; the effects of some antilipid drugs (Valeric acid, phenoxy – isobutyric acid, N-desmethyl rosuvastatin) and some antioxidant compounds (Gallic acid, Quercitin, Pyrogallol, Ascorbic acid) on the activity of immobilized hPON1 enzyme were investigated. Inhibition kinetics (IC50 and Ki values) was determined.