Purification and characterisation of glutathion s-transferase enzyme from liver tissue of Siraz (Capoeta umbla) and investigation of the effects of some chemicals on enzyme activity

Glutathione S-transferase (GST; EC 2.5.1.18) enzyme plays an important role in detoxification in metabolism. Decrease in GST enzyme activity for any reason causes detoxification mechanism to decrease. In this study, GST enzyme was purified by affinity chromatography from liver tissue of liver of Siraz fish by 55.16 times with 36.1 EU / mg protein specific activity and 73.6% yield. The purity of the purified enzyme was checked by SDS-polyacrylamide gel electrophoresis. The molecular weight was calculated to be approximately 28,6 kDa. The optimum pH of the enzyme was 7.5, the stable pH was 6,5, the optimum ionic strength was 100 mM K-phosphate and finally the optimum temperature was 35 ° C. In addition, the KM constant for the GSH substrate was 0.168 mM and the Vmax constant was 0.166 EU / ml, the KM constant for the CDNB substrate was 1.179 mM and the Vmax constant was 0.979 EU / ml. Finally, the activity of inhibition of pesticides such as esfenvalerate, deltamethrin, cypermethrin, diniconazole, and atrazine was investigated with the metal ions such as Se2-, Ba2+, Cu2+, Ag+, and Pb2+on the GST enzyme purified from liver tissue from Siraz Fish and IC50 values and Ki constants were calculated.