Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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):4677–4683
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):45–55
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram. Anal Biochem 72(1–2):248–251. https://doi.org/10.1016/0003-2697(76)90527-3
Ceyhun SB, Şentürk M, Erdoğan O, Küfrevioğlu Öİ (2010) In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills. Pestic Biochem Physiol 97(3):177–181. https://doi.org/10.1016/j.pestbp.2010.01.003
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.2011.03.013
Comakli V, Ciftci M, Kufrevioglu OI (2013) Effects of some metal ions on rainbow trout erythrocytes glutathione S-transferase enzyme: an in vitro study. J Enzyme Inhib Med Chem. 28(6):1261–1266. https://doi.org/10.3109/14756366.2012.729829
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/10.1177/0748233713475514
Çomaklı V, Kuzu M, Demirdağ R (2013) Characterization and purification of glutathione S-transferase from the liver and gill tissues of Ağrı Balık Lake trout Salmo trutta labrax and the effects of heavy metal ions on its activity. J Aquat Anim Health 27(3):145–151. https://doi.org/10.1080/08997659.2015.1032441
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.21675
Ekinci D, Beydemir S, Kufrevioglu OI (2007) In vitro inhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases. J Enzyme Inhib Med Chem 22(6):745–750. https://doi.org/10.1080/14756360601176048
Esbaugh AJ, Tufts BL (2006) The structure and function of carbonic anhydrase isozymes in the respiratory system of vertebrates. Respir Physiol Neurobiol 154(1–2):185–198. https://doi.org/10.1016/j.resp.2006.03.007
Gilmour KM, Perry SF (2009) Carbonic anhydrase and acid–base regulation in fish. J Exp Biol 212:1647–1661. https://doi.org/10.1242/jeb.029181
Hisar O, Beydemir S, Bulbul M, Yanik T (2006) Kinetic properties of carbonic anhydrase purified from gills of rainbow trout (Oncorhynchus mykiss). J Appl Anim Res 30(2):185–188. https://doi.org/10.1080/09712119.2006.9706615
Jezierska B, Ługowska K, Witeska M (2009) The effects of heavy metals on embryonic development of fish (a review). Fish Physiol Biochem 35(4):625–640. https://doi.org/10.1007/s10695-008-9284-4
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 24:429–436
Kaya ED, Söyüt H, Beydemir Ş (2013) Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: the toxicological effects of heavy metals. Environ Toxicol Pharmacol 36(2):514–521. https://doi.org/10.1016/j.etap.2013.05.019
Kaya ED, Söyüt H, Beydemir Ş (2015) The toxicological impacts of some heavy metals oncarbonic anhydrase from gilthead sea bream (Sparus aurata) gills. Environ Toxicol Pharmacol 39(2):825–832. https://doi.org/10.1016/j.etap.2015.01.021
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/s13765-015-0098-7
Kucuk M, Gulcin I (2016) Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity. Environ Toxicol Pharmacol 44:134–139. https://doi.org/10.1016/j.etap.2016.04.011
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-1404-76
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/s1069
Laemmli DK (1970) Cleavage of structural proteins during the assembly of the head. Nature 227:680–685
Manyin T, Rowe CL (2009) Bioenergetic effects of aqueous copper and cadmium the grass shrimp, Palaemonetes pugio. Comp Biochem Physiol, B 150(1):65–71. https://doi.org/10.1016/j.cbpc.2009.02.007
Mashifane TB, Moyo NAG (2014) Acute toxicity of selected heavy metals to Oreochromis mossambicus fry and fingerlings. Afr J Aquat Sci 39(3):279–285. https://doi.org/10.2989/16085914.2014.960358
Pandey N, Sharma CP (2002) Effect of heavy metals Co2+ Ni2+ and Cd2+ on growth and metabolism of cabbage. Plant Sci 163(4):753–758. https://doi.org/10.1016/S0168-9452(02)00210-8
Pane EF, Bucking C, Patel M, Wood CM (2005) Renal function in the freshwater rainbow trout (Oncorhynchus mykiss) following acute and prolonged exposure to waterborne nickel. Aquat Toxicol 72(1–2):119–133. https://doi.org/10.1016/j.aquatox.2004.11.020
Şentürk M, Gülçin İ, Beydemir Ş, Küfrevioğlu Öİ, Supuran CT (2011) In vitro inhibition of human carbonic anhydrase I and II isozymes with natural phenolic compounds. Chem Biol Drug Des 77(6):494–499. https://doi.org/10.1111/j.1747-0285.2011.01104.x
Sorsa S, Gezahagn A, Dadebo E (2016) Bioaccumulation of heavy metals in two morphotypes of African large barb Labeobarbus intermedius (Osteichthyes: Cyprinidae) in Lake Hawassa, Ethiopia. Afr J Aquat Sci 41(4):427–434. https://doi.org/10.2989/16085914.2016.1218821
Soyut H, Beydemir S (2008) Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein Pept Lett 15(5):528–535. https://doi.org/10.2174/092986608784567627
Tang CH, Lee TH (2007) The novel correlation of carbonic anhydrase II and anion exchanger 1 in gills of the spotted green pufferfish, Tetraodon nigrovirids. J Exp Zool 307A:411–418. https://doi.org/10.1002/jez.391
Teta C, Ncube M, Naik SY (2017) Heavy metal contamination of water and fish in peri-urban dams around Bulawayo, Zimbabwe. Afr J Aquat Sci 42(4):351–358. https://doi.org/10.2989/16085914.2017.1392925
Wang Q, Liu BZ, Yang HS, Wang XY, Lin ZH (2009) Toxicity of lead, cadmium and mercury on embryogenesis, survival, growth and metamorphosis of Meretrix meretrix larvae. Ecotoxicology 18(7):829–837. https://doi.org/10.1007/s10646-009-0326-1
Wilbur KM, Anderson NG (1948) Electrometric and colorimetric determination of carbonic anhydrase. J Biol Chem 176:147–154
Witeska M, Sarnowski P, Ługowska K, Kowal E (2014) The effects of cadmium and copper on embryonic and larval development of ide Leuciscus idus L. Fish Physiol Biochem 40(1):151–163. https://doi.org/10.1007/s10695-013-9832-4
Wo-Niak K, Basiak J (2003) Free radicals-mediated induction of oxidized DNA bases and DNA protein cross-links by nickel chloride. Mutat Res Genet Toxicol Environ Mutagen 514(1–2):233–243. https://doi.org/10.1016/S1383-5718(01)00344-8
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kuzu, M., Çomaklı, V., Akkemik, E. et al. Inhibitory properties of some heavy metals on carbonic anhydrase I and II isozymes activities purified from Van Lake fish (Chalcalburnus Tarichi) gill. Fish Physiol Biochem 44, 1119–1125 (2018). https://doi.org/10.1007/s10695-018-0499-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-018-0499-8