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活化能(Ea)是反应中一个重要的物理量,与反应速度直接相关,因此本实验计划利用一个新的循环催化流动分析方法(RCFA),测定愈创木酚(GA)-HRP-H2O2反应体系的活化能。RCFA系统为循环回路,人为干扰因素小,能够对酶催化过程实现连续检测,得到的测定结果准确度高。使用RCFA系统在可见光谱下测定得到GA-HRP-H2O2体系完整动力学曲线,由此求解得到酶催化反应表观速率常数(k),最后利用阿仑尼乌斯公式求得该催化反应体系的活化能(Ea)=53.46±0.097kJ.mol-1,指前因子(AE)=2.5×103±0.099min-1;Ea与AE的相对标准偏差分别为0.18%和3.81%,结果表明在一级反应阶段的不同区段中HRP催化反应的活化能和指前因子均为常数。
The activation energy (Ea) is an important physical quantity in the reaction and is directly related to the reaction rate. Therefore, this experiment plans to use a new cyclic catalytic flow analysis (RCFA) to determine the reaction system of guaiacol (GA) -HRP-H2O2 The activation energy. The RCFA system is a circulating loop with small human interference factor and can continuously detect the enzymatic catalysis process. The obtained measurement result has high accuracy. The kinetic curve of GA-HRP-H2O2 system was determined by using the RCFA system under visible light spectrum, and the apparent rate constant (k) of the enzyme-catalyzed reaction was obtained by solving the equation. Finally, the Arrhenius equation The activation energy (Ea) = 53.46 ± 0.097kJ · mol-1 and the pre-exponential factor (AE) = 2.5 × 103 ± 0.099min-1. The relative standard deviations of Ea and AE were 0.18% and 3.81%, respectively. The activation energy and the pre-exponential factor of the HRP catalysis in different sections of the reaction stage are constant.