İskorpit Balığı Solungaç Dokusundan Glutatyon Redüktaz Enziminin Saflaştırılması ve Metal İnhibisyonunun İncelenmesi

Bu çalışmada iskorpit balığı solungaç dokusundan glutatyon redüktaz enzimi literatürde ilk kez kısmi olarak saflaştırılmış ve ağır metal iyonlarının enzim aktivitesi üzerindeki etkileri belirlenmiştir. Saflaştırma işlemi homojenat hazırlanması, amonyum sülfat çöktürmesi ve diyaliz olarak üç aşamada gerçekleştirilmiştir. Çalışma sonucunda optimum pH 6.5, optimum substrat konsantrasyonu 2 mM NADPH ve optimum tampon 400 mM KH2PO4 olarak bulunmuştur. Kısmi saflaştırma sonrasında ağır metal iyonları olarak Ni+2, Zn+2 ve Cd+2’nin inhibisyon etkileri araştırılmış ve ağır metallerin IC50 değerleri sırasıyla 31 µM, 56 µM ve 74 µM olarak hesaplanmıştır. En güçlü inhibitörün çinko oldu tespit edilmiştir.

Anahtar Kelimeler:

Enzim saflaştırılması, glutatyon redüktaz, iskorpit balığı, metal iyonları

Purification and Metal Inhibition of Glutathione Reductase Enzyme from Gill Tissue of Scorpionfish

In this study, glutathione reductase enzyme was partially purified from gill tissue of scorpion fish and the effects of heavy metal ions on enzyme activity were determined. The purification process was achieved in three steps as preparation of homogenate, ammonium sulfate precipitation and dialysis. As a result of the study, optimum pH 6.5, optimum substrate concentration 2 mM NADPH and optimum buffer 400 mM KH2PO4 were determined. After partial purification, the inhibition effects of Cd+2, Ni+2, Zn+2 as heavy metal ions were investigated. The IC50 values of heavy metals were calculated as 74 µM, 31 µM, and 56 µM, respectively. The most potent inhibitor was determined to be zinc.

Keywords:

Enzyme purification, glutathione reductase, metal ion, scorpion fish,

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