Quantitative analyses of individual sugars in mixture using FRET-based biosensors = FRET 기반 바이오센서 이용 복합당 정량분석 기술

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Title
Quantitative analyses of individual sugars in mixture using FRET-based biosensors = FRET 기반 바이오센서 이용 복합당 정량분석 기술
Author(s)
J S Ha; Jongsik Gam; Su-Lim Choi; K H Oh; H S Ro; Jae Jun Song; C S Shin; Seung Goo Lee
Bibliographic Citation
Biotechnology Progress, vol. 28, no. 5, pp. 1376-1383
Publication Year
2012
Abstract
Molecular biosensors were developed and applied to measure individual sugars in biological mixtures such as bacterial culture broths. As the sensing units, four sugar-binding proteins (SBPs for allose, arabinose, ribose, and glucose) were selected from the Escherichia coli genome and connected to a cyan fluorescent protein and yellow fluorescent protein via dipeptide linkers (CFP-L-SBP-YFP). The putative sensors were randomized in the linker region (L) and then investigated with regard to the intensity of fluorescence resonance energy transfer on the binding of the respective sugars. As a result, four representatives were selected from each library and examined for their specificity using 16 available sugars. The apparent dissociation constants of the allose, arabinose, ribose, and glucose sensors were estimated to be 0.35, 0.36, 0.17, and 0.18 μM. Finally, the sugar sensors were applied to monitor the consumption rate of individual sugars in an E. coli culture broth. The individual sugar profiles exhibited a good correlation with those obtained using an HPLC method, confirming that the biosensors offer a rapid and easy-to-use method for monitoring individual sugars in mixed compositions.
Keyword
BiomassFluorescence proteinFluorescence resonance energy transferMolecular biosensorSugar
ISSN
8756-7938
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/btpr.1592
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
Synthetic Biology and Bioengineering Research Institute > 1. Journal Articles
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