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The Strathprints institutional repository is a digital archive of University of Strathclyde research outputs.

Strathprints serves world leading Open Access research by the University of Strathclyde, including research by the Strathclyde Institute of Pharmacy and Biomedical Sciences (SIPBS), where research centres such as the Industrial Biotechnology Innovation Centre (IBioIC), the Cancer Research UK Formulation Unit, SeaBioTech and the Centre for Biophotonics are based.

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Glucose sensing based on the intrinsic fluorescence of sol-gel immobilized yeast hexokinase

Hussain, F. and Birch, David J.S. and Pickup, John C. (2005) Glucose sensing based on the intrinsic fluorescence of sol-gel immobilized yeast hexokinase. Analytical Biochemistry, 339 (1). pp. 137-143. ISSN 0003-2697

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Abstract

In this study, we investigated measurements of the intrinsic fluorescence of yeast hexokinase as an assay for glucose and immobilization of the enzyme in a silica sol-get matrix as a potential in vivo glucose sensor for use in patients with diabetes. The intrinsic fluorescence of hexokinase in solution (excitation = 295 nm, emission = 330 nm) decreased by 23% at a saturating glucose concentration of 1 mM (K-d = 0.3 mM), but serum abolished the glucose-related fluorescence response. When entrapped in tetramethylorthosilicate-derived sol gel, hexokinase retained activity, with a 25% maximal glucose-related decrease in intrinsic fluorescence, and the saturation point was increased to 50 mM glucose (K-d = 12.5 mM). The glucose response range was increased further (to 120 mM, K-d = 57 mM) by a covering membrane of poly(2-hydroxyethyl) methacrylate. Unlike free enzyme, the fluorescence responses to glucose with sol-gel immobilized hexokinase, with or Without covering membrane, were similar for buffer and serum. We conclude that fluorescence monitoring of sol-gel entrapped yeast hexokinase is a Suitable system for development as an in vivo glucose biosensor. (c) 2005 Elsevier Inc. All rights reserved.