A novel and direct assay for measuring enzymatic depolymerisation of ?-1,3-glucans
| Autor: | Michael D. Brownleader, Max W. Adlard, Avni Güven, Lale Yıldız Aktaş, M Trevan |
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| Rok vydání: | 2000 |
| Předmět: |
Detection limit
chemistry.chemical_classification Chromatography biology Chemistry Plant Science General Medicine Glucanase Biochemistry In vitro Enzyme assay Analytical Chemistry Laminarin chemistry.chemical_compound Hydrolysis Enzyme Complementary and alternative medicine Drug Discovery biology.protein Molecular Medicine Quantitative analysis (chemistry) Food Science |
| Zdroj: | Phytochemical Analysis. 11:301-303 |
| ISSN: | 1099-1565 0958-0344 |
| DOI: | 10.1002/1099-1565(200009/10)11:5<301::aid-pca518>3.0.co;2-6 |
| Popis: | Indirect colorimetric analyses of released reducing groups, as an indicator of induced β-1,3-glucanase activities, tend to be the assay methods of choice for the characterization of plant endo- and exo-β-1,3-glucanase. However, interfering low molecular weight reducing sugars found in unprocessed plant extracts often mask in vitro estimations of β-1,3-glucanase activity. The enzyme-catalysed hydrolysis of an optically active β-1,3-glucan polymer was monitored polarimetrically by measuring changes in the rotation of plane-polarized light of the reaction assay medium. A direct, non-radiochemical assay that detects changes in the optical rotation of released (13) β-D-oligoglucosides with low degrees of polymerisation (≪25) has been found to be highly reproducible, rapid (total analysis time 15 min), sensitive (detection limit 0.007 unit/mL glucanase), selective and non-destructive. This assay has been used to detect a salicylate-induced cell wall-bound tomato β-1,3-glucanase, which is a component of a pathogenesis-related response in plants. Copyright © 2000 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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