title: 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.
abstract: In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77EU/mg and a yield of 35.5% using Sepharose-4B-l-tyrosine- sulphanilamide affinity column chromatography. For determining the enzyme purity and subunit molecular mass, sodiumdodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The molecular mass of subunit was found approximately 29.71kDa. The optimum temperature, activation energy (Ea), activation enthalpy (ΔH) and Q10 values were obtained from Arrhenius plot. Km and Vmax values for p-nitrophenyl acetate of the purified enzyme were calculated from Lineweaver-Burk graphs. In addition, the inhibitory effects of different heavy metal ions (Fe(2+), Pb(2+), Co(2+), Ag(+) and Cu(2+)) on Black Sea trout kidney tissue CA enzyme activities were investigated by using esterase method under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC50) were obtained. Finally Ki values and inhibition types were calculated from Lineweaver-Burk graphs.
authors.affiliation: Atatürk University, Faculty of Sciences, Department of Chemistry, Erzurum, Turkey.
authors.forename: Murat
authors.initials: M
authors.lastname: Kucuk
authors.affiliation: Atatürk University, Faculty of Sciences, Department of Chemistry, Erzurum, Turkey; King Saud University, College of Science, Department of Zoology, Riyadh, Saudi Arabia. Electronic address: igulcin@atauni.edu.tr.
authors.forename: İlhami
authors.initials: İ
authors.lastname: Gulcin
mesh_terms: D000818:Animals; D002257:Carbonic Anhydrase Inhibitors / Q000633:toxicity*; D002256:Carbonic Anhydrases / Q000302:isolation & purification / Q000378:metabolism; D029941:Fish Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D007668:Kidney / Q000201:enzymology*; D019216:Metals, Heavy / Q000633:toxicity*; D014337:Trout
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D029941:Fish Proteins; D019216:Metals, Heavy; D002256:Carbonic Anhydrases
keywords: Black Sea trout; Carbonic anhydrase; Enzyme characterization; Enzyme purification; Heavy metal; Salmo trutta Labrax