Kuzu Müslüm, Çomaklı Veysel, Akkemik Ebru, Çiftci Mehmet, Küfrevioğlu Ömer İrfan
Faculty of Pharmacy, University of Ağrı İbrahim Çeçen, 04100, Ağrı, Turkey.
School of Healthy, University of Ağrı İbrahim Çeçen, 04100, Ağrı, Turkey.
Fish Physiol Biochem. 2018 Aug;44(4):1119-1125. doi: 10.1007/s10695-018-0499-8. Epub 2018 Apr 8.
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 "CO-hydratase activity". Purity control was checked by SDS-PAGE. In vitro inhibitory effect of Cu, Ag, Cd, Ni metal ions, and arsenic (V) oxide were also examined for both isozymes activities. Whereas Cu, Ag, Cd, and Ni ions showed inhibitory effects on both isozymes, arsenic (V) oxide showed activation effect. IC values were calculated by drawing activity %-[I] graphs for metal ions exhibiting inhibitory effects. IC 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.
在本研究中，通过使用琼脂糖-4B-L-酪氨酸-磺胺亲和色谱法从凡湖鱼鳃中纯化碳酸酐酶I和II同工酶，并测定某些金属对酶活性的影响。对于纯化的碳酸酐酶I同工酶，产率、比活性和纯化倍数分别为42.07%、4948.12酶活力单位/毫克蛋白质和116.61；对于碳酸酐酶II同工酶，分别为7%、1798.56酶活力单位/毫克蛋白质和42.38。通过测量“CO水合酶活性”来测定碳酸酐酶的活性。通过SDS-PAGE检查纯度控制。还研究了铜、银、镉、镍金属离子和五氧化二砷对两种同工酶活性的体外抑制作用。铜、银、镉和镍离子对两种同工酶均表现出抑制作用，而五氧化二砷表现出激活作用。通过绘制具有抑制作用的金属离子的活性%-[I]图来计算半数抑制浓度（IC）值。碳酸酐酶I同工酶的IC值分别测定为3.39、6.38、13.52和206微摩尔，碳酸酐酶II同工酶的IC值分别为6.16、20.29、46和223微摩尔。
