Shivakumar, A. and Dinesh, R. and Krishna, H. and Nagaraja, P. (2010) Development and kinetic validation of an assay for the quantitative determination of peroxidase: Application in the detection of activity in crude plant tissues. Enzyme and Microbial Technology, 47 (6). 243 - 248. ISSN 0141-0229
Full text not available from this repository. (Request a copy)Abstract
A simple, rapid, and sensitive direct spectrophotometric assay of peroxidase activity based on its reaction with a chromogenic reagent 2,4-dimethoxyaniline (DMA) in aqueous media of pH 3.7–7.8 is reported. This method is based on the intermolecular coupling of activated DMA in the presence of peroxidase and H2O2 to produce a violet-colored species having λmax 540nm. This method has been used in the assay of H2O2 in the range of 40–330μM. At a H2O2 concentration of 660μM, the peroxidase linearity is established in the range of 0.2022–1.13 and 0.0079–0.756nM by rate and one-time detection method, respectively. Two substrate kinetic ping-pong mechanism is established. A Hanes-Woolf plot is used in the evaluation of Michaelis-Menten constants of H2O2 and dimethoxyaniline. The catalytic efficiency and catalytic power of the proposed method are 2.67×105min−1M−1 and 2.05×10−4min−1, respectively. The apparent k3 at H2O2 concentration ranging between 40 and 1320μM is found to be 469min−1. This method has been applied in the crude plant extracts with medicinal value.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Horseradish peroxidase assay, Dimethoxyaniline, Plant extracts, Catalytic parameter |
| Subjects: | C Chemical Science > Chemistry |
| Divisions: | Department of > Chemistry |
| Depositing User: | LA manjunath user |
| Date Deposited: | 05 Jul 2019 06:20 |
| Last Modified: | 11 Dec 2019 06:32 |
| URI: | http://eprints.uni-mysore.ac.in/id/eprint/4745 |
Actions (login required)
 |
View Item |
