A novel and direct assay for measuring enzymatic depolymerisation of β‐1,3‐glucans
Makaleler > Tam Makale · 2000
Yazarlar: Lale AKTAŞ, Max Adlard, Michael Trevan, Avni Guven, Michael D. Brownleader
Phytochemical Analysis, Uluslararası, Wos, 2000, cilt 11, ss 301-303
DOI: 10.1002/1099-1565(200009/10)11:5<301::aid-pca518>3.3.co;2-y
Özet
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 (1→3) 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.