A NEW FLUOROGENIC SUBSTRATE FOR HORSERADISH PEROXIDASE

摘要

Horseradish peroxidase(HRP, EC1.11.1. 7) is one of the most commonly used enzyme label for immunological [1] detection systems. It has also been used for quantitative determination of oxidases, substrates, activators, and inhibitors of such systems [2]. The interest for the development of new substrates is based on the fact that HRP decomposes two molecules of hydrogen peroxide, the natural substrate, into water and oxygen. However, the specificity of HRP for the second molecule of hydrogen peroxide is low and many other electron donors may be substituted. The aim for new substrate development was to establish a sensitive detection method for hydrogen peroxide or HRP. These substrates include chromogenic chemiluminescent or fluorogenic type. The fluorescent techniques are based on the formation of strong fluorescent oxidation products. Scopoletin [3] and diacetydichlorofluorescine [4] were first used as fluorogenic substrates. Guilbault et al. found that homovanillic acid, 4-hydroxylphenylacetate (HPA), tyrosine and tyramine could serve as fluorogenic substrates of HRP [5]. Later on, a few research groups exploited the use of other phenol derivatives [6-8].