Purification and characterization of glutathione reductase from sheep liver

ULUSU, Göktuğ; ERAT, Mustafa; Ciftci, Mehmet; ŞAKİROĞLU, HALİS; Bakan, Ebubekir

Purification and characterization of glutathione reductase from sheep liver

Göktuğ ULUSU, (Atatürk Üniversitesi, Fen Edebiyat Fakültesi, Kimya Bölümü, Erzurum, Türkiye)

Mustafa ERAT, (Atatürk Üniversitesi, Biyoteknoloji Uygulama ve Araştırma Merkezi, Erzurum, Türkiye)

Mehmet ÇİFTÇİ, (Atatürk Üniversitesi, Biyoteknoloji Uygulama ve Araştırma Merkezi, Erzurum, Türkiye)

Halis ŞAKİROĞLU, (Atatürk Üniversitesi, Fen Edebiyat Fakültesi, Kimya Bölümü, Erzurum, Türkiye)

Ebubekir BAKAN (Atatürk Üniversitesi, Fen Edebiyat Fakültesi, Biyokimya Anabilim Dalı, Erzurum, Türkiye)

Turkish Journal of Veterinary and Animal Sciences

4 0

Yıl: 2005 Cilt: 29 Sayı: 5 Sayfa Aralığı: 1109 - 1117 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Purification and characterization of glutathione reductase from sheep liver

Öz:

Glutatyon redüktaz (Glutatyon: NADP+ oksidoredüktaz, EC 1.8.1.7; GR) enzimi koyun karaciğerinden saflaştırıldı. Saflaştırma işlemi, homojenatın hazırlanması, amonyum sülfat çöktürmesi, 2', 5'-ADP Sepharose-4B affinite kromatografisi ve jel filtrasyon kromatografisi şeklinde dört basamakta yapıldı. 47,27 EÜ/mg protein spesifik aktivitesine sahip olan GR enzimi % 14,1 verimle elde edildi. Enzim için optimum pH, stabil pH, optimum sıcaklık, sırasıyla 8,0, 5,5, 60 °C olarak bulundu. Ayrıca NADPH ve GSSG substratları için $K_M$ ve $V_{max}$ değerleri sırasıyla 0,0258, 0,0239 mM ve 0,266, 0,255 EÜ/ml olarak belirlendi. Tüm saflaştırma basamakları sonucu enzim 1.477 kat saflaştırıldı ve enzim aktivitesi spektrofotometrik olarak 340 nm'de ölçüldü. Buna ilaveten NADP+ ve GSH için Lineweaver-Burk grafikleri yardımıyla K, sabitleri sırasıyla 4,367 ve 0,4055 mM olarak bulundu.

Anahtar Kelime: karaciğer pH glutatyon redüktaz (NAD(P)H) koyun enzim aktivitesi saflaştırma NADPH karakterizasyon

Konular: Ziraat Mühendisliği

Koyun karaciğerinden glutatyon redüktaz enziminin saflaştırılması ve karakterizasyonu

Öz:

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 $K_M$ ve $V_{max}$ 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 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.

Anahtar Kelime: sheep enzyme activity purification NADPH characterization liver pH glutathione reductase (NAD(P)H)

Konular: Ziraat Mühendisliği

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

TRDizin Dışındaki Atıf Sayıları

OpenCitations: 0

OpenAlex: 0

Crossref: 0

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APA	ULUSU G, ERAT M, Ciftci M, ŞAKİROĞLU H, Bakan E (2005). Purification and characterization of glutathione reductase from sheep liver. , 1109 - 1117.
Chicago	ULUSU Göktuğ,ERAT Mustafa,Ciftci Mehmet,ŞAKİROĞLU HALİS,Bakan Ebubekir Purification and characterization of glutathione reductase from sheep liver. (2005): 1109 - 1117.
MLA	ULUSU Göktuğ,ERAT Mustafa,Ciftci Mehmet,ŞAKİROĞLU HALİS,Bakan Ebubekir Purification and characterization of glutathione reductase from sheep liver. , 2005, ss.1109 - 1117.
AMA	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E Purification and characterization of glutathione reductase from sheep liver. . 2005; 1109 - 1117.
Vancouver	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E Purification and characterization of glutathione reductase from sheep liver. . 2005; 1109 - 1117.
IEEE	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E "Purification and characterization of glutathione reductase from sheep liver." , ss.1109 - 1117, 2005.
ISNAD	ULUSU, Göktuğ vd. "Purification and characterization of glutathione reductase from sheep liver". (2005), 1109-1117.

APA	ULUSU G, ERAT M, Ciftci M, ŞAKİROĞLU H, Bakan E (2005). Purification and characterization of glutathione reductase from sheep liver. Turkish Journal of Veterinary and Animal Sciences, 29(5), 1109 - 1117.
Chicago	ULUSU Göktuğ,ERAT Mustafa,Ciftci Mehmet,ŞAKİROĞLU HALİS,Bakan Ebubekir Purification and characterization of glutathione reductase from sheep liver. Turkish Journal of Veterinary and Animal Sciences 29, no.5 (2005): 1109 - 1117.
MLA	ULUSU Göktuğ,ERAT Mustafa,Ciftci Mehmet,ŞAKİROĞLU HALİS,Bakan Ebubekir Purification and characterization of glutathione reductase from sheep liver. Turkish Journal of Veterinary and Animal Sciences, vol.29, no.5, 2005, ss.1109 - 1117.
AMA	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E Purification and characterization of glutathione reductase from sheep liver. Turkish Journal of Veterinary and Animal Sciences. 2005; 29(5): 1109 - 1117.
Vancouver	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E Purification and characterization of glutathione reductase from sheep liver. Turkish Journal of Veterinary and Animal Sciences. 2005; 29(5): 1109 - 1117.
IEEE	ULUSU G,ERAT M,Ciftci M,ŞAKİROĞLU H,Bakan E "Purification and characterization of glutathione reductase from sheep liver." Turkish Journal of Veterinary and Animal Sciences, 29, ss.1109 - 1117, 2005.
ISNAD	ULUSU, Göktuğ vd. "Purification and characterization of glutathione reductase from sheep liver". Turkish Journal of Veterinary and Animal Sciences 29/5 (2005), 1109-1117.