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木瓜蛋白酶具有过氧化物模拟酶活性,可以催化过氧化氢氧化3,3’,5,5’-四甲基联苯胺(TMB)产生蓝色的显色反应。结合尿酸酶和木瓜蛋白酶构建了一个比色检测尿酸(UA)的新方法。其检测过程受pH、温度、TMB浓度和反应时间的影响,最优的检测条件分别为pH 3.0、温度为30℃、TMB浓度0.80 mmol/L及反应时间2 h,线性范围为20~200μmol/L,检出限2.4μmol/L。实验证实了该方法具有较好的选择性并且已经成功地应用于人血清样品的尿酸测定。
Papain has a peroxidase mimic enzyme activity that catalyzes the oxidation of 3,3 ’, 5,5’-tetramethylbenzidine (TMB) by hydrogen peroxide to produce a blue color reaction. A new method of colorimetric detection of uric acid (UA) was constructed by combining uricase and papain. The optimum conditions were pH 3.0, temperature 30 ℃, TMB concentration 0.80 mmol / L and reaction time 2 h, and the linear range was 20 ~ 200 μmol / L. The detection conditions were pH, temperature, TMB concentration and reaction time. L, the detection limit of 2.4μmol / L. Experiments show that the method has good selectivity and has been successfully applied to the determination of uric acid in human serum samples.