title: Orthogonal Surface Tags for Whole-Cell Biocatalysis.
abstract: We herein describe the engineering of E. coli strains that display orthogonal tags for immobilization on their surface and overexpress a functional heterologous "protein content" in their cytosol at the same time. Using the outer membrane protein Lpp-ompA, cell-surface display of the streptavidin-binding peptide, the SpyTag/SpyCatcher system, or a HaloTag variant allowed us to generate bacterial strains that can selectively bind to solid substrates, as demonstrated with magnetic microbeads. The simultaneous cytosolic expression of functional content was demonstrated for fluorescent proteins or stereoselective ketoreductase enzymes. The latter strains gave high selectivities for specific immobilization onto complementary surfaces and also in the whole-cell stereospecific transformation of a prochiral CS-symmetric nitrodiketone.
authors.affiliation: Karlsruhe Institute of Technology (KIT), Institute for Biological Interfaces (IBG 1), Hermann-von-Helmholtz-Platz, 76344, Eggenstein-Leopoldshafen, Germany.
authors.forename: Theo
authors.initials: T
authors.lastname: Peschke
authors.affiliation: Karlsruhe Institute of Technology (KIT), Institute for Biological Interfaces (IBG 1), Hermann-von-Helmholtz-Platz, 76344, Eggenstein-Leopoldshafen, Germany.
authors.forename: Kersten S
authors.initials: KS
authors.lastname: Rabe
authors.affiliation: Karlsruhe Institute of Technology (KIT), Institute for Biological Interfaces (IBG 1), Hermann-von-Helmholtz-Platz, 76344, Eggenstein-Leopoldshafen, Germany.
authors.forename: Christof M
authors.initials: CM
authors.lastname: Niemeyer
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D001665:Binding Sites; D055162:Biocatalysis; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D018914:Cells, Immobilized / Q000166:cytology / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D008074:Lipoproteins / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D013237:Stereoisomerism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D008074:Lipoproteins; C099932:Lpp protein, E coli; D008164:Luminescent Proteins; C452700:streptavidin-binding peptide; C054562:yellow fluorescent protein, Bacteria; C078167:OMPA outer membrane proteins
keywords: bacteria; biocatalysis; cell-surface display; immobilization techniques; stereoselective reactions