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Characterization and inhibition effects of some metal ions on carbonic anhydrase enzyme from Kangal Akkaraman sheep
Author(s) -
Kocyigit Umit M.,
Taslimi Parham,
Gulçin İlhami
Publication year - 2018
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.22172
Subject(s) - chemistry , carbonic anhydrase , enzyme , chromatography , metal ions in aqueous solution , molecular mass , esterase , metal , enzyme assay , gel electrophoresis , affinity chromatography , polyacrylamide gel electrophoresis , substrate (aquarium) , yield (engineering) , carbonic anhydrase i , sodium , sodium dodecyl sulfate , nuclear chemistry , biochemistry , organic chemistry , biology , ecology , materials science , metallurgy
In this work, the carbonic anhydrase (CA) enzyme was purified from Kangal Akkaraman sheep in Sivas, Turkey with specific activity value of 6681.57 EU/mg and yield of 14.90% with using affinity column chromatography. For designating the subunit molecular mass and enzyme purity, sodium dodecyl sulfate polyacrylamide gel electrophoresis method was used and single band for this procedure was obtained. The molecular mass of CA enzyme was found as 28.89 kDa. In this study, the optimum temperature and optimum pH were obtained from 30 and 7.5. V max and K m values for p‐nitrophenylacetate substrate of the CA were determined from Lineweaver–Burk graphs. Additionally, the inhibitory results of diverse heavy metal ions (Hg + , Fe 2+ , Pb 2+ , Co 2+ , Ag + , and Cu 2+ ) on sheep were studied. Indeed, CA enzyme activities of Kangal sheep were investigated with using esterase procedure under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC 50 ) and K i values were obtained.