Glutatyon S-Transferaz Enziminin Tavuk Karaciğerinden Saflaştırılması ve Karakterizasyonu

Abstract

In this study, the glutathione S-transferase enzyme (GST; EC 2.5.1.18) was purified with 8.35 EU/mL specific activity, 24.56 times 8% yield, from chicken liver, using ammonium sulfate precipitation and glutathione-agarose affinity chromatography. In order to control the purity of the enzyme, SDS-PAGE was performed and a single band was obtained. The molecular mass of the subunit was calculated as approximately 30.9 kDa. In addition, the optimum pH value of the enzyme (8.5 in Tris-HCl); optimum ionic strength (150 mM with Tris-HCl); optimum temperature (70 oC); stable pH value (8.5 with Tris-HCl) was determined. The KM value for the GSH substrate of the enzyme was 0.802 mM, the Vmax value was 1.833 EU/mL; For CDNB, the KM value was calculated as 3.6 mM and the Vmax value was calculated as 2.829 EU/mL.