Rapid and direct spectrophotometric method for kinetics studies and routine assay of peroxidase based on aniline diazo substrates
| Autor: | Fatemeh Mirazizi, Azita Bahrami, Mehdi Bakavoli, Hossein Shahbani Zahiri, Raymond L. Legge, Kamahldin Haghbeen |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Aniline Compounds
Kinetics 010501 environmental sciences 01 natural sciences Substrate Specificity chemistry.chemical_compound Aniline Spectrophotometry Drug Discovery medicine Organic chemistry 0105 earth and related environmental sciences Peroxidase Pharmacology Detection limit Chromatography medicine.diagnostic_test biology Spectrum Analysis 010401 analytical chemistry Substrate (chemistry) General Medicine 0104 chemical sciences chemistry biology.protein Diazo Azo Compounds |
| Zdroj: | Journal of enzyme inhibition and medicinal chemistry. 31(6) |
| ISSN: | 1475-6374 |
| Popis: | Peroxidases are ubiquitous enzymes that play an important role in living organisms. Current spectrophotometrically based peroxidase assay methods are based on the production of chromophoric substances at the end of the enzymatic reaction. The ambiguity regarding the formation and identity of the final chromophoric product and its possible reactions with other molecules have raised concerns about the accuracy of these methods. This can be of serious concern in inhibition studies. A novel spectrophotometric assay for peroxidase, based on direct measurement of a soluble aniline diazo substrate, is introduced. In addition to the routine assays, this method can be used in comprehensive kinetics studies. 4-[(4-Sulfophenyl)azo]aniline (λmax = 390 nm, ɛ = 32 880 M(-1) cm(-1) at pH 4.5 to 9) was introduced for routine assay of peroxidase. This compound is commercially available and is indexed as a food dye. Using this method, a detection limit of 0.05 nmol mL(-1) was achieved for peroxidase. |
| Databáze: | OpenAIRE |
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