title: Quantitative analyses of individual sugars in mixture using FRET-based biosensors.
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.
authors: Systems & Synthetic Biology Research Center, KRIBB, Daejeon 305-806, Korea.
authors: Jae-Seok
authors: JS
authors: Ha
authors: Jongsik
authors: J
authors: Gam
authors: Su-Lim
authors: SL
authors: Choi
authors: Ki-Hoon
authors: KH
authors: Oh
authors: Hyeon-Su
authors: HS
authors: Ro
authors: Jae Jun
authors: JJ
authors: Song
authors: Chul Soo
authors: CS
authors: Shin
authors: Seung-Goo
authors: SG
authors: Lee
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000032:analysis*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D007700:Kinetics
chemical_list: D002241:Carbohydrates