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Dergiler / Turkish Journal of Veterinary and Animal Sciences / 2005 / Cilt: 29 - Sayı: 5

Purification and Characterization of Glutathione Reductase from Sheep Liver

Göktuğ ULUSU
Sayfa
1109–1117
Yayın Tarihi
2005-05-01
DOI
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In addition, Ki values of 4.367 and 0.4055 mM were determined by means of Lineweaver-Burk graphs for NADP+ and GSH, respectively.

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Amaç & Hipotez:Glutathione reductase (Glutathione: NADP+ oxidoreductase, EC 1.8.1.7; GR) was purified from sheep liver.
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Temel Bulgular & Veri:The purification included 4 steps: preparation of homogenate, ammonium sulfate fractionation, 2', 5'-ADP Sepharose-4B affinity chromatography, and gel filtration chromatography.
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Sonuç & Katkı:In addition, Ki values of 4.367 and 0.4055 mM were determined by means of Lineweaver-Burk graphs for NADP+ and GSH, respectively.
🔬Tür:Araştırma Makalesi
📍Kapsam:Genişletilmiş Akademik Örneklem
⚙️Değişkenler:Glutathione reductase · purification · characterization

Özet

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. KM and Vmax 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 nm. In addition, Ki values of 4.367 and 0.4055 mM were determined by means of Lineweaver-Burk graphs for NADP+ and GSH, respectively.

Abstract

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. KM and Vmax 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 nm. In addition, Ki values of 4.367 and 0.4055 mM were determined by means of Lineweaver-Burk graphs for NADP+ and GSH, respectively.

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Bu Makaleye Atıf Yap

ULUSU, G. (2005). Purification and Characterization of Glutathione Reductase from Sheep Liver. Turkish Journal of Veterinary and Animal Sciences, Cilt: 29 - Sayı: 5, 1109–1117. https://ajindex.com/articles/122423

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Kaynakça (9)

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