title: Utilization of organophosphate:phosphate antiporter for isotope-labeling experiments in E. coli.
abstract: The transport of organophosphates across the cytoplasma membrane is mediated by organophosphate:phosphate antiporter proteins. In this work, we present the application of a recombinant phosphoenolpyruvate:phosphate antiporter for isotopic labeling experiments in E. coli strains. The antiporters UhpT, UhpT-D388C, and PgtP were investigated regarding transport activity and growth on phosphoenolpyruvate as sole carbon source. The expression of the protein variant UhpT-D388C in a shikimic acid producing E. coli strain was used to show the successful isotopic labeling of shikimic acid from extracellular phosphoenolpyruvate. The results demonstrate the possibility of a direct incorporation of exogenously applicated glycolysis intermediates into E. coli cells for13C-labeling experiments.
authors.affiliation: Institute of Microbiology, Universität Stuttgart, Stuttgart, Germany christoph.albermann@imb.uni-stuttgart.de.
authors.forename: Christoph
authors.initials: C
authors.lastname: Albermann
authors.affiliation: Institute of Biochemical Engineering, Technische Universität München, Garching, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Weiner
authors.affiliation: Institute of Biochemical Engineering, Technische Universität München, Garching, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Tröndle
authors.affiliation: Institute of Biochemical Engineering, Technische Universität München, Garching, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Weuster-Botz
authors.affiliation: Institute of Microbiology, Universität Stuttgart, Stuttgart, Germany.
authors.forename: Georg A
authors.initials: GA
authors.lastname: Sprenger
mesh_terms: D017920:Antiporters / Q000235:genetics / Q000378:metabolism*; D002247:Carbon Isotopes / Q000032:analysis; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D007553:Isotope Labeling; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010755:Organophosphates / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism*; D012765:Shikimic Acid / Q000378:metabolism*
chemical_list: D017920:Antiporters; D002247:Carbon Isotopes; D029968:Escherichia coli Proteins; D009004:Monosaccharide Transport Proteins; D010755:Organophosphates; D010710:Phosphates; C063498:UhpT protein, E coli; D012765:Shikimic Acid
keywords: isotopic labeling; organophosphate:phosphate antiporter; phosphoenolpyruvate; shikimic acid