斑马鱼尿酸酶基本特性研究

The Study on the Properties of Uricase from Zebrafish

分别研究了反应环境、处理环境对斑马鱼尿酸酶活力以及稳定性的影响。在不同温度及酸碱度环境下分别测定斑马鱼尿酸酶的活性来确定其最适反应温度、pH值。通过测定在不同温度及酸碱度条件下处理后斑马鱼尿酸酶的残余活力来确定其稳定性。通过测定不同pH值缓冲体系下制备的斑马鱼尿酸酶活力来确定其最佳溶解pH值。用Lineweaver-Burk作图法计算斑马鱼尿酸酶的Km值。斑马鱼尿酸酶最佳溶解pH值为8.0～9.5。其最适反应温度为40～45℃。其最适反应pH值为8.4～9.0。其温度在50℃以下、pH值为7.0～9.0时稳定性最好。另外该酶与尿酸底物反应过程的米氏常数(Km)为18.67μmol/L。

To investigate the effects of reaction condition on the uricase activity of zebrafish, its thermal stability, pH-stabil-ity and other basic features.Authors mensurated the enzyme activity under the different temperature, different pH to confirm its optimum reaction condition, thermal stability, pH stability and optimum dissolution.At last, authors used the constructing method of Lineweaver Burk to confirm the Km of uricase.According to the results, the optimum dissolution pH value was range from 8.0 to 9.5.On the stated condition, the range of optimum pH and range of pH stability were 8.0～9.0 and 7.0～9.0.The optimal reaction temperature was between 40～45℃ and showed good thermal stability with below 50℃.Finally, the Km of purified uricase was 18.67 μmol/L at pH 8.4 and 37℃.