In this study, CA I and II isoenzymes were purified from Van Lake fish gills by using Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography and to determine the effects of some metals on the enzyme activities. For purified CA I isoenzyme, yield, specific activity, and purification fold were obtained as 42.07%, 4948.12 EU/mg protein, and 116.61 and for CA II isoenzyme, 7%, 1798.56 EU/mg protein, and 42.38 respectively. Activity of CA was determined by measuring “CO2-hydratase activity”. Purity control was checked by SDS-PAGE. In vitro inhibitory effect of Cu2+, Ag+, Cd2+, Ni2+ metal ions, and arsenic (V) oxide were also examined for both isozymes activities. Whereas Cu2+, Ag+, Cd2+, and Ni2+ ions showed inhibitory effects on both isozymes, arsenic (V) oxide showed activation effect. IC50 values were calculated by drawing activity %-[I] graphs for metal ions exhibiting inhibitory effects. IC50 values were determined as 3.39, 6.38, 13.52, and 206 μM for CA I isozyme and 6.16, 20.29, 46, and 223 μM for CA II isozyme respectively.
Carbonic anhydrase Heavy metal Inhibition Purification
This is a preview of subscription content, log in to check access.
Akyol H, Kuzu M (2017) In vitro effects of some heavy metal ions on cytosolic thioredoxin reductase purified from rainbow trout gill tissues. Fresenius Environ Bull 26(7):4677–4683Google Scholar
Beydemir Ş, Bülbül M, Hisar O, Söyüt H, Yanık T (2006) Carbonic anhydrase: affinity purification and kinetic properties from rainbow trout lens. IJAC 2(1):45–55Google Scholar
Ceyhun SB, Sentürk M, Yerlikaya E, Erdogan O, Küfrevioglu OI, Ekinci D (2011) Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity. Environ Toxicol Pharmacol 32(1):69–74. https://doi.org/10.1016/j.etap.2011.03.013CrossRefPubMedGoogle Scholar
Comakli V, Akkemik E, Ciftci M, Küfrevioğlu Öİ (2015) Purification and characterization of glucose 6-phosphate dehydrogenase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the effects of some metals on enzyme activity. Toxicol Ind Health 1(5):403–411. https://doi.org/10.1177/0748233713475514CrossRefGoogle Scholar
Demirdag R, Comakli V, Kuzu M, Yerlikaya E, Şentürk M (2015) Purification and characterization of carbonic anhydrase from Ağrı Balık Lake Trout Gill (Salmo trutta labrax) and effects of sulfonamides on enzyme activity. J Biochem Mol Toxicol 29(3):123–128. https://doi.org/10.1002/jbt.21675CrossRefPubMedGoogle Scholar
Kalay M, Canlı M (2000) Elimination of essential (Cu, Zn) and nonessential (Cd, Pb) metals from tissues of a freshwater fish Tilapia zillii following an uptake protocol. Turk J Zool 24:429–436Google Scholar
Kho KH, Kim JW, Kim SC, Choi MR, Han KH, Lee WK, Choi KS (2015) Identification and intracellular localization of carbonic anhydrase I in gills, heart, muscle, and intestine of rainbow trout, Oncorhynchus mykiss. J Korean Soc Appl Biol Chem 58(5):729–733. https://doi.org/10.1007/s13765-015-0098-7CrossRefGoogle Scholar
Kuzu M, Ciftci M (2015) Purification and characterization of NADPH-cytochrome P450 reductase from Lake Van fish liver microsomes and investigation of some chemical and metals’ effects on the enzyme activity. Turk J Chem 39(1):149–158. https://doi.org/10.3906/kim-1404-76CrossRefGoogle Scholar
Kuzu M, Aslan A, Ahmed I, Comakli V, Demirdag R, Uzun N (2016) Purification of glucose-6-phosphate dehydrogenase and glutathione reductase enzymes from the gill tissue of Lake Van fish and analyzing the effects of some chalcone derivatives on enzyme activities. Fish Physiol Biochem 42(2):483–491. https://doi.org/10.1007/s1069CrossRefPubMedGoogle Scholar