基于辣根过氧化物酶催化H2O2氧化邻苯二胺新体系的反应动力学研究

[The study on reaction kinetics based on a new system of the horseradish peroxidase catalyting the oxidation of o-phenylenediamine by H2O2].

作者信息

Jiang Yuan-liang, Feng Chun-liang

机构信息

Department of Chemistry, Liaoning Normal University, Dalian 116029, China.

出版信息

Guang Pu Xue Yu Guang Pu Fen Xi. 2002 Jun;22(3):436-40.

PMID:12938327

Abstract

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 horseradish 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 nm, pH = 4.85, t = 22 degrees C 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(-3) mol.L-1 and upsilon m = 7.2 x 10(-2) delta A.min-1.

摘要

本文研究了辣根过氧化物酶催化H2O2氧化邻苯二胺合成2,3-二氨基吩嗪化合物的酶催化反应动力学。首先，合成了H2O2在辣根过氧化物酶（HRP）催化下氧化邻苯二胺（OPD）的氧化产物。选择了最佳反应条件（酸度、温度、OPD、H2O2、HRP的各种摩尔比）。通过紫外吸收光谱、红外光谱和元素分析确定了反应条件和产物。结果表明产物为2,3-二氨基吩嗪。其次，基于米氏方程，采用紫外分光光度法研究了OPD-H2O2-HRP体系的反应级数和速率常数。选择了最佳条件λmax = 446.3 nm，pH = 4.85，t = 22℃，进行了一系列实验。实验结果表明，该反应经实验和理论确定为单反应物反应，得出Km = 6.02×10-3 mol·L-1，vm = 7.2×10-2 ΔA·min-1。