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- Title
Kinetic Study of PeroxidaseCatalyzed Coupling of Benzene1,4diamine and N2Aminoethylnaphthalen1amine: Development of Micromolar Hydrogen Peroxide Reaction System.
- Authors
Anantharaman Shivakumar; Padmarajaiah Nagaraja; Narayanan Aradhana; AvinashK. Gowda
- Abstract
A novel chromogenic method to measure the peroxidase activity using paraphenylenediamine dihydrochloride =benzene1,4diamine hydrochloride; PPDD and N1naphthylethylenediamine dihydrochloride =N2aminoethylnaphthalen1amine; NEDA is presented. The PPDD entraps the free radical and gets oxidized to electrophilic diimine, which couples with NEDA to give an intense redcolored chromogenic species with maximum absorbance at 490nm. This assay was adopted for the quantification of H2O2between 20 and 160μM. Catalytic efficiency and catalytic power of the commercial peroxidase were found to be 4.47×104M−1min−1and 3.38×10−4min−1, respectively. The catalytic constant kcat and specificity constant kcatKm at saturated concentration of the cosubstrates were 0.0245×103min−1and 0.0445μM−1min−1, respectively. The chromogenic coupling reaction has a minimum interference from the reducing substances such as ascorbic acid, Lcystein, citric acid, and oxalic acid. The method being simple, rapid, precise, and sensitive, its applicability has been tested in the crude vegetable extracts that showed peroxidase activity.
- Subjects
ENZYME kinetics; PEROXIDASE; BENZENE; AMINES; NAPHTHALENE; HYDROGEN peroxide; NITROGEN; ELECTROPHILES
- Publication
Helvetica Chimica Acta, 2010, Vol 93, Issue 6, p1227
- ISSN
0018-019X
- Publication type
Article