title: Escherichia [corrected] coli ribose binding protein based bioreporters revisited.
abstract: Bioreporter bacteria, i.e., strains engineered to respond to chemical exposure by production of reporter proteins, have attracted wide interest because of their potential to offer cheap and simple alternative analytics for specified compounds or conditions. Bioreporter construction has mostly exploited the natural variation of sensory proteins, but it has been proposed that computational design of new substrate binding properties could lead to completely novel detection specificities at very low affinities. Here we reconstruct a bioreporter system based on the native Escherichia coli ribose binding protein RbsB and one of its computationally designed variants, reported to be capable of binding 2,4,6-trinitrotoluene (TNT). Our results show in vivo reporter induction at 50 nM ribose, and a 125 nM affinity constant for in vitro ribose binding to RbsB. In contrast, the purified published TNT-binding variant did not bind TNT nor did TNT cause induction of the E. coli reporter system.
authors.affiliation: Department of Fundamental Microbiology, University of Lausanne, Bâtiment Biophore, Quartier UNIL-Sorge 1015 Lausanne, Switzerland.
authors.forename: Artur
authors.initials: A
authors.lastname: Reimer
authors.affiliation: Department of Plant and Environmental Sciences, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
authors.forename: Sharon
authors.initials: S
authors.lastname: Yagur-Kroll
authors.affiliation: Department of Plant and Environmental Sciences, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
authors.forename: Shimshon
authors.initials: S
authors.lastname: Belkin
authors.affiliation: Department of Fundamental Microbiology, University of Lausanne, Bâtiment Biophore, Quartier UNIL-Sorge 1015 Lausanne, Switzerland.
authors.forename: Shantanu
authors.initials: S
authors.lastname: Roy
authors.affiliation: Department of Fundamental Microbiology, University of Lausanne, Bâtiment Biophore, Quartier UNIL-Sorge 1015 Lausanne, Switzerland.
authors.forename: Jan Roelof
authors.initials: JR
authors.lastname: van der Meer
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods; D002352:Carrier Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D033902:Periplasmic Binding Proteins / Q000378:metabolism*; D012266:Ribose / Q000378:metabolism*; D014303:Trinitrotoluene / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D033902:Periplasmic Binding Proteins; C028003:RbsB protein, E coli; D014303:Trinitrotoluene; D012266:Ribose