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Clé, Carla, Gunning, A. Patrick, Syson, Karl, Bowater, Laura, Field, Robert A. 
ORCID: https://orcid.org/0000-0001-8574-0275 and Bornemann, Stephen
(2008)
Detection of transglucosidase-catalyzed polysaccharide synthesis on a surface in real time using surface plasmon resonance spectroscopy.
Journal of the American Chemical Society, 130 (46).
pp. 15234-15235.
ISSN 0002-7863
Abstract
Biological systems that involve enzyme catalysis at surfaces, particularly strategically important ones that involve insoluble substrates/products such as the cell wall and the starch granule, require analyses beyond classical solution state enzymology. Using a model system, we have demonstrated the real-time measurement of transglucosidase activity on a surface using surface plasmon resonance (SPR) spectroscopy. We monitored the extension of a (partially carboxymethylated) dextran surface with alternansucrase and sucrose as a glycosyl donor. Conditions were used where surface polymer synthesis rates were a function of enzyme concentration and proportional to the extent of enzyme binding to the surface. A method to determine the turnover number of the enzyme on the surface was also developed. The presence of a new amorphous polysaccharide was observed optically, detected by lectin binding and imaged by atomic force microscopy. This surface method will have utility in a wide range of carbohydrate enzyme systems including screens.
| Item Type: | Article |
|---|---|
| Faculty \ School: | Faculty of Medicine and Health Sciences > Norwich Medical School Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemistry, Pharmacy and Pharmacology Faculty of Science > School of Chemical Sciences and Pharmacy (former - to 2009) |
| Depositing User: | Rhiannon Harvey |
| Date Deposited: | 31 May 2011 10:58 |
| Last Modified: | 24 Sep 2024 09:31 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/31555 |
| DOI: | 10.1021/ja805264w |
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