Vocational School of Health Services, Cumhuriyet University, Sivas, 58140, Turkey.
Department of Chemistry, Faculty of Science, Atatürk University, Erzurum, 25240, Turkey.
J Biochem Mol Toxicol. 2018 Aug;32(8):e22172. doi: 10.1002/jbt.22172. Epub 2018 Jul 9.
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 and K 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 , Pb , Co , Ag , and Cu ) 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 ) and K values were obtained.
在这项工作中，使用亲和层析柱从土耳其锡瓦斯的 Kangal Akkaraman 绵羊中纯化了碳酸酐酶 (CA)，其比活值为 6681.57 EU/mg，得率为 14.90%。为了指定亚基分子量和酶纯度，使用了十二烷基硫酸钠聚丙烯酰胺凝胶电泳法，得到了单一的条带。CA 酶的分子量为 28.89 kDa。在这项研究中，最适温度和最适 pH 值分别为 30 和 7.5。通过 Lineweaver-Burk 图确定了 CA 对 p-硝基苯乙酸酯底物的 V 和 K 值。此外，还研究了不同重金属离子 (Hg、Fe、Pb、Co、Ag 和 Cu) 对绵羊的抑制作用。事实上，在体外条件下，使用酯酶法研究了 Kangal 绵羊的 CA 酶活性。获得了抑制 50%酶活性的重金属浓度（IC ）和 K 值。
