Glucose-sensing proteins from mesophilic and thermophilic bacteria as new tools in diabetes monitoring

S. D'Auria; Mose Rossi; Joseph R. Lakowicz

doi:10.1117/12.426739

10 May 2001 Glucose-sensing proteins from mesophilic and thermophilic bacteria as new tools in diabetes monitoring

S. D'Auria, Mose Rossi, Joseph R. Lakowicz

Proceedings Volume 4252, Advances in Fluorescence Sensing Technology V; (2001) https://doi.org/10.1117/12.426739
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

We developed a new method of glucose sensing using inactive forms of glucose oxidase from Aspergillus niger and glucose dehydrogenase from the thermophilic microorganism Thermoplasma acidophilum. Glucose oxidase was rendered inactive by removal of the FAD cofactor. The resulting apo- glucose oxidase still binds glucose as observed from a decrease in its intrinsic tryptophan fluorescence. 8- Anilino-1-naphthalene sulfonic acid (ANS) was found to bind spontaneously to apo-glucose oxidase as seen from an enhancement of the ANS fluorescence. The steady state intensity of the bound ANS decreased 25% upon binding of glucose, and the mean lifetime of the bound ANS decreased about 40%. These spectral changes occurred with a midpoint from 10 to 20 mM glucose, which is comparable to the K_o of holo-glucose oxidase. These results suggest that apo- glucose oxidase can be used as a reversible non-consuming sensor for glucose.

Citation Download Citation

S. D'Auria, Mose Rossi, and Joseph R. Lakowicz "Glucose-sensing proteins from mesophilic and thermophilic bacteria as new tools in diabetes monitoring", Proc. SPIE 4252, Advances in Fluorescence Sensing Technology V, (10 May 2001); https://doi.org/10.1117/12.426739

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