SavedAdd your journal

Journals / Uluslararası Gıda Tarım ve Hayvan Bilimleri Dergisi / 2026 / Cilt: 6 Sayı: 2

Purification and Characterization of Glutathione Reductase Enzyme from Sheep Heart

Glutatyon Redüktaz Enziminin Koyun Kalp Dokusundan Saflaştırılması ve Karakterizasyonu

Cansu Bulak(BİNGÖL ÜNİVERSİTESİ)0009-0002-8939-2567
Yusuf Temel(BİNGÖL ÜNİVERSİTESİ)0000-0001-8148-3718
Mehmet Çiftci(BİNGÖL ÜNİVERSİTESİ)0000-0002-1748-3729
Pages
29–43
Publication Date
2026-09-29
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

In addition, the KM constant for NADPH was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL; and the KM constant for GSSG was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Glutathione reductase (GR; E.C.1.8.1.7) is an enzyme that catalyzes the reduction of GSSG to GSH and is a type of oxidoreductase.
📊
Key Findings & Data:Purification was carried out using 30-70% salt precipitation and affinity chromatography.
💡
Conclusion & Impact:In addition, the KM constant for NADPH was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL; and the KM constant for GSSG was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL.
🔬Design:Araştırma Makalesi
📍Scope:Comprehensive Sample
⚙️Variables:Glutatyon Redüktaz · Saflaştırma · Karakterizasyon

Abstract

Glutathione reductase (GR; E.C.1.8.1.7) is an enzyme that catalyzes the reduction of GSSG to GSH and is a type of oxidoreductase. In this study, glutathione reductase was purified from sheep heart tissue. Purification was carried out using 30-70% salt precipitation and affinity chromatography. The enzyme, with a specific activity of 5.35 EU/mg protein, was purified 191-fold at 72.82%. The purification status of the enzyme was then checked by electrophoresis, and its molecular mass was found to be approximately 70 kDa under denaturing conditions. In characterization studies, the enzyme's optimum pH was found to be 6.0 (in 0.1 M KH2PO4 buffer), its stable pH was found to be 8.0 (in 0.1 M KH2PO4 buffer), its optimum ionic strength was found to be 240 mM (0.1 M KH2PO4), and its optimum temperature was found to be 70°C. In addition, the KM constant for NADPH was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL; and the KM constant for GSSG was found to be 7.776 mM, and the Vmax value was found to be 0.1678 EU/mL.

Özet

Glutatyon redüktaz (GR; E.C.1.8.1.7), GSSG’nin GSH’a indirgenmesini katalizleyen bir enzim olup oksidoredüktaz sınıfının bir çeşididir. Bu çalışmada glutatyon redüktaz enzimi koyun kalp dokusundan saflaştırıldı. Saflaştırma işleminde %30-70 arası tuz çöktürmesi ve afinite kromatografisi metodları kullanıldı. Saflaştırma sonucunda spesifik aktivitesi 5,35 EÜ/mg.protein olan enzim %72,82 oranında 191 kat şeklinde saflaştırıldı. Daha sonra enzimin saflaştırılma durumu elektroforez ile kontrol edildi ve denatüre edici şartlarda molekül kütlesi yaklaşık olarak 70 kDa olarak bulundu. Karakterizasyon çalışmalarında ise enzimin optimum pH’sı 6,0 (0,1 M KH2PO4 tamponunda), stabil pH’sı 8,0 (0,1 M KH2PO4 tamponunda), optimum iyonik şiddetti 240 mM (0,1 M KH2PO4) ve optimum sıcaklık değeri 70°C olarak bulundu. Ayrıca NADPH için KM sabiti 7,776 mM, Vmax değeri 0,1678 EÜ/mL; GSSG için KM sabiti 7,776 mM, Vmax değeri 0,1678 EÜ/mL olarak tespit edildi.

🔖

Cite This Article

Bulak, C., Temel, Y., Çiftci, M. (2026). Purification and Characterization of Glutathione Reductase Enzyme from Sheep Heart. Uluslararası Gıda Tarım ve Hayvan Bilimleri Dergisi, Cilt: 6 Sayı: 2, 29–43. https://ajindex.com/articles/1623425

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (23)

  1. Acan, N. L., Tezcan, E. F. (1989). Sheep brain glutathione reductase: purification and general porperties. FEBS Letter, 250 (1), 72-74.
  2. Aksoy, M., Kıranşan, K. D. (2020). The constructon and testing of an amperometric biosensor for oxidized glutathione with glutathione reductase ımmobilized on reduced graphene oxide paper modified with cobalt sulphur. Chemistry Select., 5, 13925-13935.
  3. Barea, J. L., Lee, C. Y. (1979). Mouse liver glutathione reductase purification, kinetics and regulation. Eur. J. Biochem., 98, 487.499.
  4. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principple of protein-dye binding. Analytical Biochemistry, 72, 248-254.
  5. Can, B., Kulaksız Erkmen, G., Dalmızrak, Ö., Ogus, I.H., Özer, N. (2010). Purification and characterization of rat kidney glutathione reductase. Protein J., 29, 250-256.
  6. Carlberg, I., Mannervik, B. (1975). Purfication and charracterization of the glutathione reductase from rat liver. The Journal of Biologıcal Chemistry, 250(14), 5475-5480.
  7. Carlberg, I., Mannervik, B. (1985). Reduction of 2,4,6-Trinitrobenzenesulfonate by glutathione reductase and effect of NADP⁺ on the electron transfer. The Journal of Biologıcal Chemistry, 261 (4), 1629-1635.
  8. Demir, D., Yörük, İ. H. (2022). Diplotaenia turcia bitkisinde glutatyon redüktaz ve glutatyon s-transferaz enzim aktiviteleri ile protein karbonil düzeyinin belirlenmesi. BÜSAD, 3(2), 165-172.
  9. Dudak, S. B., Işık, K., Mitrovica B., Soydan, E., Ekinci, D. (2023). Purification of glutathione reductase enzyme from whiting fish gill tissue and investıgatıon of metal inhibition. Open Access Journal, 6(6), 610-614.
  10. Erat, M., Demir, H., Şakiroğlu, H. (2005). Purification of glutathione reductase from chicken liver and ınvesgation of kinetic properties. Applied biochemistry and biotechnology, 125, 127-138.
  11. Erat, M., Şakiroğlu, H., Çiftci, M. (2003). Purification and characterization Of glutathione reductase from bovine erythrocytes. Preparative Biochemistry & Biotechnology, 33(4), 283-300.
  12. Gutteridge, J. M. C. (1995). Lipid peroxidation and antioxidants as biomarkers of damage. Clin. Chem., 41 (12), 1819-1828.
  13. Karadağ, H. (2019). The in vitro effect of neonicotinoid insecticides imidacloprid and thiamethoxam on glutathione reductase enzyme activity from Baker’s Yeast (Saccharomyces Crevisia). ADYUJSCI, 9 (2), 266-273.
  14. Laemmli, D. K. (1970). Cleavage of srructual proteins during in aaaembly of the heat of bacteriaophage T4. Nature, 227, 680.
  15. Lineweaver, H., Burk, D. (1934). The determination of enzyme dissociation constants. J. Am. Chem. Soc., 57, 658-666.
  16. Sies, H. (1991). Oxidative stress from basic reseachto clinical application. Am. J. Med., 91, 31-38.
  17. Tandoğan, B., Ulusu, N. N. (2006). Kinetic mechanism and molecular properties of glutathione reductase. FABAD J. Pharm. Sci., 31, 230-237.
  18. Tandoğan, B., Ulusu, N. N. (2010). Purification and kinetics of bovine kidney cortex glutahione reductase. Protein &Peptide Letters, 17, 667-674.
  19. Taşer, P., Çiftci, M. (2012). Purification and characterization of glutathione reductase from turkey liver. Turkish Journal of Veterinary& Animal Sciences, 36(5), 546-553.
  20. Temel, Y., Taher, S. S. M., Hamza, M. A., Shafeeq, İ. H., Koçyiğit, U. M. ve Çiftci, M. (2018) Bazı metal iyonlarının japon bıldırcın (Coturnix Coturnix Japonica) karaciğer glutatyon redüktaz enzimi aktivitesi üzerine inhibisyon etkilerinin araştırılması. Cumhuriyet Scie.J., 39(3), 679-687.
  21. Ulusu, G., Erat, M., Çiftci, M., Şakiroğlu, H., Bakan, E. (2003). Purification and characterization of glutathione reductase from sheep liver. Turk. J. Vet. Anim. Sci. 29, 1109-1117.
  22. Ulusu, N. N., Tandoğan, B. (2007). Purification and kinetic of glutathione reductase from bovine liver. Mol. Cell. Biochem., 303, 45-51.
  23. Voskava, J., Kocan, L., Vasko, L. and Perjesi, P. (2023). Glutathione related enzymes and proteins: a review. Molecules, 28, 1447.