EFFECTS OF FERULIC ACID ON HUMAN SERUM PARAOXONASE ENZYME PURIFIED BY THREE PHASE PARTITIONING

Yıl 2021, Cilt: 1 Sayı: 1, 1 - 7, 30.12.2021

Semra Çiçek

Öz

Human serum paraoxonase 1 (PON1) is a enzyme which inhibits macrophage cholesterol biosynthesis, metabolizes peroxides of cholesterol esters and reduces cholesterol efflux into macrophages. Therefore, it is speculated to play a role in several human diseases including diabetes mellitus and atherosclerosis. In this study, paraoxonase enzyme was first purified from human serum with three-phase partitioning method (TPP), also kinetic, characterization studies, and effects of ferulic acid were carried out. TPP purification process was performed in three stages. In first stage, PON1 was exposed to 60-80% ammonium sulfate precipitation, in the second stage, 1,0:0,5 ratio of human serum/ t-butanol and 20% ammonium sulfate saturation were used. In the third stage, the second TPP stage made over the intermediate, PON1 enzyme was purified from human serum with 49,87% recovery and 182,66 purification fold using constant ratio of human serum:t-butanol. In studies of the purified enzyme for kinetic properties, optimum pH, stable pH, optimum temperature, stable temperature were determined as 8,1, 7,0, 37°C, 20°C, respectively. Molecular weight of enzyme was found to be 45 kDa from Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). In addition, the inhibition effects of ferulic acid on the purified PON1 enzyme were investigated and Lineweaver Burk plots were obtained.

Anahtar Kelimeler

Paraoxonase 1, Three-Phase Partitioning Method, Ferulic Acid

Kaynakça

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