title: Analysis of Enantioselective Biotransformations Using a Few Hundred Cells on an Integrated Microfluidic Chip.
abstract: The investigation of stereoselective biocatalytic transformations at a single-cell level is to date an unsolved challenge. Here, we report the development of an integrated microfluidic device which enables the analytical characterization of enantioselective reactions at nanoliter scale by combining whole-cell catalyzed on-chip syntheses, chiral microchip electrophoresis, and label-free detection of enantiomers by deep UV time-resolved fluorescence. Using Escherichia coli expressing recombinant Aspergillus niger epoxide hydrolase as the model enzyme for various enantioselective reactions, we evaluated the approach for downscaling the reaction to a few hundred cells. Our work is thus an important step toward the analysis of single-cell stereoselective biocatalysis.
authors.affiliation: Institute of Analytical Chemistry, University of Leipzig , Linnéstrasse 3, 04103 Leipzig, Germany.
authors.forename: Karin M
authors.initials: KM
authors.lastname: Krone
authors.affiliation: Institute of Analytical Chemistry, University of Leipzig , Linnéstrasse 3, 04103 Leipzig, Germany.
authors.forename: Rico
authors.initials: R
authors.lastname: Warias
authors.affiliation: Faculty of Chemistry, Philipps-Universität Marburg , Hans-Meerwein-Strasse, 35032 Marburg, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Ritter
authors.affiliation: Faculty of Chemistry, Philipps-Universität Marburg , Hans-Meerwein-Strasse, 35032 Marburg, Germany.|Max-Planck-Institut für Kohlenforschung , Kaiser-Wilhelm-Platz 1, 45470 Mülheim/Ruhr, Germany.
authors.forename: Aitao
authors.initials: A
authors.lastname: Li
authors.affiliation: Faculty of Chemistry, Philipps-Universität Marburg , Hans-Meerwein-Strasse, 35032 Marburg, Germany.|Max-Planck-Institut für Kohlenforschung , Kaiser-Wilhelm-Platz 1, 45470 Mülheim/Ruhr, Germany.
authors.forename: Carlos G
authors.initials: CG
authors.lastname: Acevedo-Rocha
authors.affiliation: Faculty of Chemistry, Philipps-Universität Marburg , Hans-Meerwein-Strasse, 35032 Marburg, Germany.|Max-Planck-Institut für Kohlenforschung , Kaiser-Wilhelm-Platz 1, 45470 Mülheim/Ruhr, Germany.
authors.forename: Manfred T
authors.initials: MT
authors.lastname: Reetz
authors.affiliation: Institute of Analytical Chemistry, University of Leipzig , Linnéstrasse 3, 04103 Leipzig, Germany.
authors.forename: Detlev
authors.initials: D
authors.lastname: Belder
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology; D055162:Biocatalysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism*; D046210:Microfluidic Analytical Techniques; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis; D013237:Stereoisomerism
chemical_list: D011994:Recombinant Proteins; D006867:Hydrolases