Carbonic Anyhdrase Enzyme From The Siirt Mohair Goat Liver : Purification, Characterization and Assessment of Enzyme Kinetics Against Metal Toxicity

Abstract

Because of their physiological and clinical roles, carbonic anhydrases CAs are the most stud- ied enzymes. In earlier studies; CA enzymes have been purified and characterized from the tissues and erythrocytes of many organisms such as; dog, swine, sheep, chicken, bee, fish, bovine, bacteria and human. In this study, the CA enzyme has purified from Siirt Mohair Goat liver tissue with 1930.84 EU x mg-1 of specific activity, 57.28% of purification yield and 80.55 of purification folds. The purity of the purified enzyme has confirmed by SDS-PAGE. As the characterization of CA enzyme’s in Siirt Mohair Goat liver has been done; the optimum ionic strength=25 mM, the optimum pH= 8.0, the optimum temperature= 40°C and the stable pH= 7.0 has been determined. Inhibitory effects of some metal ions have been examined on the purified CA enzyme. IC50 values of inhibiting metal ions were found as 2.24, 2.76, 2.36, 3.20, 2.55, 2.25, 3.28, 2.13, 3.10, 1.75, 2.16 and 3.50 mM for Al3+, Ni2+, Cd2+, Cu2+, Pb2+, Ba2+, Zn2+, B3+, Fe3+, Se2+, Ag+ and Co2+ respectively. As a result, CA enzyme was first purified from the Siirt Mohair Goat liver and the characteristics of the enzyme were investigated in this study.

Keywords

• Affinity,metal ions,Inhibition,Siirt Mohair Goat

Karbonik Anhidraz Enziminin Siirt Tiftik Keçisi Karaciğerinden Saflaştırılması, Karakterizasyonu ve Metal Toksisitesine Karşı Enzim Kinetiğinin Değerlendirilmesi

Abstract

F izyolojik ve klinik özelliklerinden dolayı karbonik anhidrazlar CAs , en çok incelenen enzimlerdir. Daha önceki çalışmalarda CA enzimleri köpek, domuz, koyun, tavuk, arı, balık, sığır, bakteri ve insan gibi birçok canlının dokularından ve eritrositlerinden saflaştırılmış ve karakterize edilmiştir. Bu çalışmada da CA enzimi Siirt Tiftik Keçisi karaciğer dokusundan 1930.84 EU.mg-1 spesifik aktivite ile %57.28 verimle 80.55 kat saflaştırıldı. Saflaştırılan enzimin saflığı SDS-PAGE ile doğrulandı. Siirt Tiftik Keçisi karaciğerinden CA enziminin karakterizasyonu yapıldığında; optimum iyonik şiddet: 25 mM, optimum pH: 8.0, optimum sıcaklık: 40ºC ve stabil pH: 7.0 olarak belirlendi. Saflaştırılmış CA enzimi üzerine bazı metal iyonlarının inhibitör etkileri incelendi. İnhibisyon gösteren metal iyonlarının IC50 değerleri Al+3, Ni+2, Cd+2, Cu+2, Pb+2, Ba+2, Zn+2, B+3, Fe+3, Se+2, Ag+ and Co+2 için sırasıyla 2.24, 2.76, 2.36, 3.20, 2.55, 2.25, 3.28, 2.13, 3.10, 1.75, 2.16 ve 3.50 mM olarak bulundu. Sonuç olarak bu çalışmada, CA enzimi Siirt Tiftik Keçisi karaciğerinden ilk defa saflaştırıldı ve enzimin karakteristik özellikleri incelendi

Keywords

• Afinite,metal iyonları,inhibisyon,Siirt tiftik keçisi

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