| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1388213 | Carbohydrate Research | 2012 | 5 Pages |
A chromatographic method using HPAEC-PAD was developed to accurately quantify the major oligosaccharides derived from lichenase degradation of barley β-glucan. This method was further used to follow β-glucosidase degradation and product formation as progress curves. This approach allowed us to compare the kinetic characteristics of β-glucosidase on each exclusive oligosaccharide substrate and their mixtures. Our results show that when determining the kinetic parameters of exohydrolases on oligosaccharides following the progress curve for the substrates is necessary since calculations based only on released monosaccharide products may lead to an error. The catalytic activity of almond β-glucosidase on laminaribiose (G3G) was approximately half that measured against cellobiose (3.15 S−1). The enzyme had 12 times and 560 times less catalytic activity on 3-O-β-cellobiosyl-d-glucose (G4G3G) and on 3-O-β-cellotriosyl-d-glucose (G4G4G3G) respectively than on G3G. Our approach offers a useful tool for the determination of the kinetics of enzymatic or chemical modification of various carbohydrate substrates.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Major oligosaccharides released by lichenase from barely β-glucan quantified. ► Kinetics of β-glucosidase on these oligosaccharides simultaneously studied. ► Approach developed for kinetic characterization of carbohydrate substrates.
