title: Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer.
abstract: Recent improvements in Förster resonance energy transfer (FRET) sensors have enabled their use to detect various small molecules including ions and amino acids. However, the innate weak signal intensity of FRET sensors is a major challenge that prevents their application in various fields and makes the use of expensive, high-end fluorometers necessary. Previously, we built a cost-effective, high-performance FRET analyzer that can specifically measure the ratio of two emission wavelength bands (530 and 480 nm) to achieve high detection sensitivity. More recently, it was discovered that FRET sensors with bacterial periplasmic binding proteins detect ligands with maximum sensitivity in the critical temperature range of 50 - 55 °C. This report describes a protocol for assessing sugar content in commercially-available beverage samples using our portable FRET analyzer with a temperature-specific FRET sensor. Our results showed that the additional preheating process of the FRET sensor significantly increases the FRET ratio signal, to enable more accurate measurement of sugar content. The custom-made FRET analyzer and sensor were successfully applied to quantify the sugar content in three types of commercial beverages. We anticipate that further size reduction and performance enhancement of the equipment will facilitate the use of hand-held analyzers in environments where high-end equipment is not available.
authors: Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology.
authors: Haseong
authors: H
authors: Kim
authors: Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology.
authors: Gui Hwan
authors: GH
authors: Han
authors: Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology.
authors: Yaoyao
authors: Y
authors: Fu
authors: College of Interdisciplinary & Creative Studies, Konyang University.
authors: Jongsik
authors: J
authors: Gam
authors: Synthetic Biology & Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology; Biosystems and Bioengineering Program, University of Science and Technology; sglee@kribb.re.kr.
authors: Seung Goo
authors: SG
authors: Lee
mesh_terms: D001628:Beverages; D002241:Carbohydrates; D031541:Fluorescence Resonance Energy Transfer; D008024:Ligands; D033902:Periplasmic Binding Proteins; D012680:Sensitivity and Specificity
chemical_list: D002241:Carbohydrates; D008024:Ligands; D033902:Periplasmic Binding Proteins