1H NMR Spectroscopy-Based Configurational Analysis of Mono- and Disaccharides and Detection of β-Glucosidase Activity: An Undergraduate Biochemistry Laboratory
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[1] Eastern Illinois University
Eastern Illinois University
Township of Charleston, Estados Unidos
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- Localización: Journal of chemical education, ISSN 0021-9584, Vol. 92, Nº 7, 2015, págs. 1244-1249
- Idioma: inglés
- Texto completo no disponible (Saber más ...)
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Resumen
A 1H NMR spectroscopy-based laboratory experiment explores mono- and disaccharide structural chemistry, and the enzyme-substrate specificity of glycosidic bond cleavage by β-glucosidase towards cellobiose (β-linked gluco-disaccharide) and maltose (α-linked gluco-disaccharide). Structural differences between cellobiose, maltose, and the glycolytic product glucose are established in terms of chemical shift changes and J-coupling constants of the anomeric proton peaks. The distinct anomeric proton signals observed in the 4.0–5.5 ppm range of glucose, cellobiose, and maltose spectra serve as a signature NMR feature to monitor the substrate specificity of β-glucosidase towards cellobiose as a substrate. This experiment consolidates carbohydrate structural chemistry and NMR spectroscopy as applied to investigate enzyme specificity in the context of a biochemistry laboratory experiment with hands-on NMR experience for upper-division undergraduates.
