Namık KILINÇ, Zuhal ALIM, Mehmet Mustafa İŞGÖR, Şükrü BEYDEMİR

Bazı Metallerin Minekop Umbrina Cirrosa Solungacı Karbonik Anhidraz Aktivitesi Üzerine Etkileri

S Metal toksisitesi balıklarda oksidatif strese sebep olur. Bu durum insanlar ve diğer balık tüketen canlılar için potansiyel bir risk faktörüdür. Bu çalışmada minekop balığı solungacından saflaştırılan karbonik anhidraz enzimi üzerine bazı ağır metallerin inhibisyon etkileri incelendi. Minekop balığı solungacından, karbonik anhidraz, Sepharose 4B–L-tyrosin sulfanilamit afinite kolonu kullanılarak 2093,9 EUmg−1 spesifik aktiviteyle ve 86,51% verimle yaklaşık 160 kat saflaştırıldı. SDS-poliakrilamid jel elektroforezinde tek band gözlendi ve buradan enzimin molekül kütlesi yaklaşık olarak 30,8 kDa olarak belirlendi. Bazı ağır metallerin Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Ni2+ CA enzimi aktivitesi üzerine etkileri hidrataz metodu kullanılarak in vitro olarak incelendi. Sonuç olarak, ağır metallerin inhibisyon etkileri Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Ni2+ şeklinde belirlendi. Bu sonuçlar Ag+’nın en önemli potansiyel CA inhibitörü olduğunu gösterdi

Anahtar Kelimeler:

Karbonik anhidraz, solungaç, ağır metaller, inhibisyon

The impacts of some metals on the activity of corb gill Umbrina cirrosa Carbonic Anhydrase

Metal toxicity causes oxidative stress in fish. This situation is a potential risk factor for humans and other living feeding on contaminated fish. In this study, the inhibition effects of heavy metals on carbonic anhy- drase enzyme from the corb fish gill were investigated. The carbonic anhydrase enzyme was purified from gill of corb fish with a specific activity of 2093,9 EUmg−1 and 86,51% yield and approximately 160 fold using Sep- harose 4B–L-tyrosine sulfanilamide affinity chromatography method. SDS–polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approximately 30,8 kDa. Inhibitory effects of metals Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Ni2+ on CA activity were determined at different concentrations using the hydratase method under in vitro conditions. Consequently, in vitro inhibition rank order was determined as Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Ni2+. From these results, we showed that Ag+ is the most potent inhibitor of CA enzyme.

Keywords:

Carbonic anhydrase, Gill, Heavy metals, Inhibition,

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