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Purification And Characterization Of Glutathione Reductase From Sheep Liver, Göktuğ Ulusu, Mustafa Erat, Mehmet Çi̇ftçi̇, Hali̇s Şaki̇roğlu, Ebubeki̇r Bakan
Purification And Characterization Of Glutathione Reductase From Sheep Liver, Göktuğ Ulusu, Mustafa Erat, Mehmet Çi̇ftçi̇, Hali̇s Şaki̇roğlu, Ebubeki̇r Bakan
Turkish Journal of Veterinary & Animal Sciences
Glutathione reductase (Glutathione: NADP^+ oxidoreductase, EC 1.8.1.7; GR) was purified from sheep liver. The purification included 4 steps: preparation of homogenate, ammonium sulfate fractionation, 2', 5'-ADP Sepharose-4B affinity chromatography, and gel filtration chromatography. GR was obtained with a yield of 14.1% having a specific activity of 47.27 EU/mg proteins. Optimal pH, stable pH, and optimal temperature were 8.0, 5.5, and 60 °C, respectively. K_M and V_max values for NADPH and GSSG substrates were 0.0258 and 0.0239 mM, and 0.266 and 0.255 EU/ml, respectively. The overall purification was about 1477-fold for liver GR. The enzyme activity was measured spectrophotometrically at 340 …