期刊
ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY
卷 44, 期 -, 页码 134-139出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.etap.2016.04.011
关键词
Black Sea trout; Salmo trutta Labrax; Enzyme purification; Enzyme characterization; Carbonic anhydrase; Heavy metal
In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77 EU/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 29.71 kDa. The optimum temperature, activation energy (E-a), activation enthalpy (Delta H) and Q(10) values were obtained from Arrhenius plot. K-m and V-max 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 (Fe2+, Pb2+, Co2+, Ag+ and Cu2+) 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 K-i values and inhibition types were calculated from Lineweaver-Burk graphs. (C) 2016 Elsevier B.V. All rights reserved.
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