Studies on kinetic method for detecting active intermediate in enzyme reaction

A new kinetic method is proposed for detecting active intermediate in enzyme reaction, Due to the reactivity of the intermediate, a proper competition reagent which reacts with the intermediate while the intermediate changes to final product, can be found and added. By detecting the kinetic process of intermediate reaction with reagent, the information of intermediate can be obtained. The method can be used to study free radical intermediate and non-free-radical intermediate, to study fast reaction and to follow transient process, With this method, the intermediate process in oxidation reaction of norepinephrine with hydrogen peroxide, catalyzed by horseradish peroxidase, is investigated.