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Orthogonal Surface Tags for Whole‐Cell Biocatalysis
Author(s) -
Peschke Theo,
Rabe Kersten S.,
Niemeyer Christof M.
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201609590
Subject(s) - biocatalysis , cytosol , streptavidin , heterologous , chemistry , stereospecificity , cell , peptide , enzyme , biochemistry , membrane , transformation (genetics) , protein engineering , stereoselectivity , combinatorial chemistry , gene , biotin , ionic liquid , catalysis
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 C S ‐symmetric nitrodiketone.