Purifcation and Characterization of Glutathion Reductase Enzyme From Japanese Quail (Coturnix coturnix japanica) Erythrocytes

Year 2017, Volume: 7 Issue: 3, 143 - 150, 30.09.2017

Yusuf Temel Taner Bozkuş Yusuf Karagözoğlu Mehmet Çiftçi

Abstract

Glutathione (γ-L-glutamyl-L-cysteinyl-glycine) is a low molecular weight thiol which protects
organism from harmful effect of oxidized molecules in cell. Glutathione reductase (GR) is a key enzyme involved
in the glutathione metabolism. In the present study GR enzyme has been purifed and characterized from Japanese
quail erythrocytes. The purifcation proses consisted of two steps, which include preparation of hemolysate, and
2’,5’-ADP Sepharose 4B afnity chromatography. The japanese quail erythrocytes GR enzyme had specifc activity
of 33.75 EU m g-1 proteins, with a yield of 46.2% and 1 028 purifcation fold. Enzyme purity and molecular weight
of subunits were determined by SDS-PAGE method as 78.7 kDa. Then, characteristics and kinetic properties of the
purifed enzyme was determined.

Keywords

Characterization, erythrocyte, glutathion reductase, purifcation, quail

Glutatyon Redüktaz (GR) Enziminin Japon Bıldırcın (Coturnix coturnix japanica) Eritrositlerinden Saflaştırılması ve Karakterizasyonu

Abstract

Glutatyon (γ-L-glutamil-L-cysteinyl-glycine) organizmanın hücre içi okside moleküllerin zararlı
etkisinden korunmasında görevli düşük molekül ağırlıklı bir tiyoldür. Glutatyon redüktaz (GR) enzimi glutatyon
metabolizmasında yer alan temel enzimdir. Bu çalışma kapsamında GR enzimi japon bıldırcın eritrositlerinden
saflaştırılarak karakterize edilmiştir. Saflaştırma işlemi hemolizatın hazırlanması ve 2’,5’-ADP Sepharose 4B
afnite kromatografsi olmak üzere 2 basamakta gerçekleştirilmiştir. GR enzimi bıldırcın eritrositlerinden 33.75
EÜ m g-1 protein spesifk aktivite ile %46.2 verimle, 1 028 kat saflaştırılmıştır. Enzim saflığı SDS-PAGE ile
kontrol edilmiştir alt birimlerinin molekül ağırlığı aynı yöntemle 78.7 kDa olarak bulunmuştur. Daha sonra çalışma
kapsamında saflaştırılan enzimin karakteristik ve kinetik özellikleri tesbit edilmiştir.

Keywords

Bıldırcın, eritrosit, glutatyon redüktaz, karakterizasyon, saflaştırma

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