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Turkish Journal of Fisheries and Aquatic Sciences 2016, Vol 16, Num, 1     (Pages: 169-175)

Purification of Carbonic Anhydrase from Capoeta umbla (Heckel, 1843) Gills and Toxicological Effects of Some Metals on Enzyme Activity

Muammer Kırıcı 1 ,Mahinur Kırıcı 1 ,Şükrü Beydemir 3 ,Muhammed Atamanalp 4

1 Bingöl University, Faculty of Agriculture, Department of Fisheries, 12000-Bingöl, Turkey
2 Bingöl University, Faculty of Arts and Science, Department of Chemistry, 12000-Bingöl, Turkey
3 Atatürk University, Faculty of Science, Department of Chemistry, 25240-Erzurum, Turkey
4 Atatürk University, Faculty of Fisheries, 25240-Erzurum, Turkey
DOI : 10.4194/1303-2712-v16_1_17 Viewed : 3959 - Downloaded : 3071 In this study, in vitroeffects of some metal ions(Fe3+, Cd2+, Pb2+ and Ni2+)on cytoplasmic carbonic anhydrase(CA, EC from Capoeta umbla gill was investigated. CA was purified from the gills of C. umbla for the first time. It was purified with the Sepharose-4B-L-Tyrosine Sulphanilamide affinity chromatography method. The overall purification was approx. 31.69-fold with a yield of 53.33%, and a specific activity of 326.73 EU/mg proteins. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) showed a single band corresponding to a molecular weight of approx. 29 kDa. The constants of the enzyme inhibitor complex (Ki) and 50% inhibitory values (IC50) for metal ions were determined by Lineweaver-Burk graphs and plotting activity % vs. [I], respectively. The Kiconstants and IC50 values were 0.012±0.0135 and 0.136 mM for Fe3+, 0.019±0.0113 and 0.191 mM for Cd2+, 0.041±0.0075 and 0.289 mM for Pb2+, and 0,120±0.034 and 0.924 mM for Ni2+. It was determined that Fe3+, Cd2+ and Pb2+ inhibited the enzyme competitively while Ni2+ inhibited the enzyme noncompetitively. The potential inhibitor for C. umbla gill CA was found as Fe3+ from these results. Keywords : Capoeta umbla, carbonic anhydrase, gills, metal toxicity