以铍试剂Ⅱ为底物电化学法测定辣根过氧化物酶

Electrochemical Detection of Horseradish Peroxidase with BerryllonⅡ as a New Substrate

提出了以铍试剂 作为辣根过氧化物酶 ( HRP)底物伏安法测定辣根过氧化物酶活性的新方法。铍试剂 本身为偶氮结构 ,具有电化学活性 ,能够在汞电极上发生还原反应而产生一个灵敏的二阶导数还原峰 ;以 H2 O2 为氧化剂 ,加入 HRP催化 H2 O2 氧化铍试剂 反应的进行 ,使底物被氧化分解 ,其平衡浓度降低 ,对应的还原峰电流降低 ,峰电流的降低值同 HRP的加入量在 8.0× 1 0 - 7～ 8.0× 1 0 - 6 g·m L- 1之间呈线性关系 ;

A new electrochemical substrate berryllonⅡ for horseradish peroxidase (HRP) was studied for the detection of HRP activity for the first time. BerryllonⅡ can be oxidized by H 2O 2 and the addition of HRP can greatly increase the decolorizing reaction processes. It can also be easily reduced on the mercury drops electrode and a well-defined polarographic peak occurs at -360 mV. After the addition of HRP, the decrease of the peak current is in proportion to the concentration of HRP in the range of 8.0×10 -7 ～8.0×10 -6 g·mL -1 . The conditions for enzymatic reaction and voltammetric detection are optimized and the mechanism of oxidative reaction is discussed primarily.