辣根过氧化物酶催化过氧化氢氧化邻苯二胺反应动力学的研究

The Study on Reaction Kinetics Based on a New System of the Horseradish Peroxidase Catalyting the Oxidation of o-phenylenediamine by H_2O_2

本文对辣根过氧化物酶催化过氧化氢与氧化邻苯二胺合成 2 ,3 二氨基吩嗪反应动力学进行了研究。首先是用辣根过氧化物酶催化过氧化氢氧化邻苯二胺合成了氧化产物 ,并且对反应条件、酸度 ,温度 ,邻苯二胺 ,过氧化氢与辣根过氧化物酶不同的摩尔比等进行了研究。用紫外 可见光谱、红外光谱以及元素分析法对该产物进行了表征 ,结果表明 ,其产物为 2 ,3 二氨基吩嗪。然后用紫外可见分光光度法 ,根据Michaelis Menten(M M )方程研究了辣根过氧化物酶催化过氧化化氢氧化邻苯二胺反应 ,选择了最佳实验条件 ,确定了最大吸收波长λ =4 4 6 3nm ,pH =4 85和反应温度t=2 2℃ ,进行了一系列实验 ,从而确定了体系的反应级数 ,其结果表明 ,这个反应可以视为单底物反应进行处理 ,并且在实验和理论方面作出了论证 ,并且还求出Km =6 0 2× 10 - 3mol·L- 1 ,vm =7 2× 10 - 2 ΔA·min- 1 。

In this paper, we study the enzyme catalysis reaction kinetics that 2,3-diaminophenazine compound was synthesized with the horseradish peroxidase catalyzing reaction of H2O2 oxidizing o-phenylemediamine. First, the product of oxidation of o-phenylenediamine (OPD) by H2O2 catalyzed and by hotseradish peroxidase (HRP) was synthesized. The finest reaction conditions (acid degree,temperature,OPD, H2O2, HRP various molar ratio) were selected. The reaction conditions and the product were determined by UV-absorption spectrum, IR and element analysis. The results showed that the product is 2,3-diaminophenazine. Secondly, the reaction order of OPD-H2O2-HRP system and the rate constant were studied by using ultraviolet spectrophotometry based on Michaelis-Menten equation. The finest conditions lambda(max) = 446.3 =,pH = 4.85, t = 22degreesC were selected,a series experiments were done. Experimental results showed that this is an one reactant reaction determined by the experiment and theory, and derived K-m 6.02 x 10(-1) mol . L-1 and nu(m) = 7.2 x 10(-2) DeltaA-min(-1).