The correlation between some metal concentrations and carbonic anhydrase activity in Tuna Thunnus Thynnus Linnaeus, 1758 Gill

Abstract

Today metal pollution is rapidly increasing with advances in technology and industry. Due to their bioaccumulative and nonbiodegradable properties, heavy metals are important contaminant of aquatic organism all over the world. Metal toxicity causes oxidative damage such as DNA damage, enhanced lipid peroxidation, the oxidation of protein sulfydryl groups and enzyme inactivation in the metabolism. In this study, we investigated in vitro effects of some metals on carbonic anhydrase enzyme which has vital role in fish metabolism. For this aim, The enzyme, was purified from gills of tuna with a specific activity of 1062 EU/mg proteins and 31% yield using Sepharose 4B– aniline–sulfanilamide affinity chromatography method. SDS–polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approximately 29 kDa. Inhibitory effects of metals Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Co2+ 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+> Co2+. From these results, we showed that Ag+ is the most potent inhibitor of CA enzyme.

Keywords

• Carbonic anhydrase,metal toxicity,Inhibition

Orkinos Thunnus Thynnus Linnaeus, 1758 solungacında karbonik anhidraz enzim aktivitesi ve bazı metal derişimleri arasındaki ilişki

Abstract

B ugün teknoloji ve sanayideki gelişmelerle birlikte metal kirliliği hızla artmaktadır. Ağır metaller biyobirikimleri ve doğada çözülmeyen özelliklerinden dolayı tüm dünyada sucul organizma için önemli kirliliktir. Metal toksisitesi, metabolizmada DNA hasarı, lipid peroksidasyonunun artması, protein sulfidril gruplarının oksidasyonu ve enzim inaktivasyonu gibi oksidatif hasara neden olur. Bu çalışmada, balık metabolizmasında hayati rol oynayan karbonik anhidraz enzimi üzerine bazı metallerin in vitro etkilerini inceledik. Bu amaçla enzim orkinos solungaçlarından Sepharose 4B–anilin–sulfanilamid afinite kromatografisi yöntemi kullanılarak 1062 EÜ/mg protein spesifik aktivite ve %31 verimle saflaştırıldı. SDS-PAGE jel elektroforezinde yaklaşık 29 kDa’da tek bant görüldü. İn vitro koşullar altında hidrataz yöntemi kullanılarak metallerin Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Co2+ farklı derişimlerinde inhibisyon etkileri belirlendi. İn vitro inhibisyon derecesi Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Co2+ olarak belirlendi. Bu sonuçlardan Ag+’nin güçlü bir CA enzim inhibitörü olduğu görüldü

Keywords

• Karbonik anhidraz,metal toksisitesi,inhibisyon

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