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Proceedings Paper

FRET-based glucose monitoring for bioprocessing
Author(s): Amelita Bartolome; Lauren Smalls-Mantey; Debora Lin; Govind Rao; Leah Tolosa
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Paper Abstract

The glucose-mediated conformational changes in the glucose binding protein (GBP) have been exploited in the development of fluorescence based glucose sensors. The fluorescence response is generated by a polarity sensitive dye attached to a specific site. Such fluorescent sensors respond to submicromolar glucose at diffusion-controlled rates mimicking the wild type. However, such sensors have been limited to in vitro glucose sensing because of the preliminary dye-labeling step. In the study described here, the dye-labeling step is omitted by genetically encoding the GBP with two green fluorescent mutants namely, the green fluorescent protein (GFP) and the yellow fluorescent protein (YFP) in the N- and C-terminal ends, respectively. These two GFP mutants comprise a fluorescence resonance energy transfer (FRET) donor and acceptor pair. Thus, when glucose binds with GBP, the conformational changes affect the FRET efficiency yielding a dose-dependent response. A potential application for this FRET-based glucose biosensor is online glucose sensing in bioprocessing and cell culture. This was demonstrated by the measurement of glucose consumption in yeast fermentation. Further development of this system should yield in vivo measurement of glucose in bioprocesses.

Paper Details

Date Published: 23 February 2006
PDF: 8 pages
Proc. SPIE 6098, Genetically Engineered Probes for Biomedical Applications, 60980L (23 February 2006); doi: 10.1117/12.646799
Show Author Affiliations
Amelita Bartolome, Univ. of Maryland, Baltimore County (United States)
Lauren Smalls-Mantey, Univ. of Maryland, Baltimore County (United States)
Debora Lin, Univ. of Maryland, Baltimore County (United States)
Govind Rao, Univ. of Maryland, Baltimore County (United States)
Leah Tolosa, Univ. of Maryland, Baltimore County (United States)

Published in SPIE Proceedings Vol. 6098:
Genetically Engineered Probes for Biomedical Applications
Alexander P. Savitsky; Rebekka M. Wachter, Editor(s)

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