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In vitro enzymatic response of Turkish native chicken “Gerze” to heavy metal exposure

机译:土耳其土鸡“ Gerze”对重金属暴露的体外酶促反应

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Abstract Glucose-6-phosphate dehydrogenase (G6PD) was purified and characterized from the Turkish native chicken, Gerze, erythrocytes for the first time, and some characteristics were investigated. Purification procedure consisted of ammonium sulphate fractionation and affinity chromatography on 29, 59-ADP Sepharose-4B. The enzyme was purified 1063.22-fold with a yield of 43.27% and specific activity of 93.5 EU/mg proteins. Kinetic parameters of the enzyme were determined with glucose-6-phosphate (G6P) as substrate and purified enzyme had an apparent KM and Vmax values of 0.222?mM and 0.097?U/ml, respectively. The same parameters were determined with NADP+ and the KM and Vmax values were 0.0603?mM and 0.153?U/ml, respectively. The following metals, Cd+2, Pb+2, Hg+2, Cu+2, Zn+2 and Fe+3 showed inhibitory effects on the enzyme. Cd+2 and Pb+2 exhibited the strongest inhibitory action. Hg+2 and Cu+2 were moderate inhibitors, whereas Zn+2 and Fe+3 showed weaker actions. All tested metals inhibited the enzyme in competitive manner.
机译:摘要首次从土耳其土生鸡Gerze,红细胞中纯化了6-葡萄糖葡萄糖脱氢酶(G6PD),并对其特性进行了研究。纯化程序包括硫酸铵分级分离和在29,59-ADP Sepharose-4B上的亲和色谱。将该酶纯化1063.22倍,产率为43.27%,比活性为93.5EU / mg蛋白。以6-磷酸葡萄糖(G6P)为底物测定酶的动力学参数,纯化后的酶的表观K M 和V max 值为0.222?mM和0.097。 ΔU/ ml。用NADP + 测定相同的参数,K M 和V max 值分别为0.0603?mM和0.153?U / ml。下列金属Cd +2 ,Pb +2 ,Hg +2 ,Cu +2 ,Zn < sup> +2 和Fe +3 对酶具有抑制作用。 Cd +2 和Pb +2 表现出最强的抑制作用。 Hg +2 和Cu +2 是中度抑制剂,而Zn +2 和Fe +3 的作用较弱。 。所有测试的金属都以竞争方式抑制了酶。

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