Directed Evolution of an Artificial Imine Reductase

Martina Hestericová, Tillman Heinisch, Lur Alonso-Cotchico, Jean Didier Maréchal, Pietro Vidossich, Thomas R. Ward

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36 Citations (Scopus)


© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Artificial metalloenzymes, resulting from incorporation of a metal cofactor within a host protein, have received increasing attention in the last decade. The directed evolution is presented of an artificial transfer hydrogenase (ATHase) based on the biotin-streptavidin technology using a straightforward procedure allowing screening in cell-free extracts. Two streptavidin isoforms were yielded with improved catalytic activity and selectivity for the reduction of cyclic imines. The evolved ATHases were stable under biphasic catalytic conditions. The X-ray structure analysis reveals that introducing bulky residues within the active site results in flexibility changes of the cofactor, thus increasing exposure of the metal to the protein surface and leading to a reversal of enantioselectivity. This hypothesis was confirmed by a multiscale approach based mostly on molecular dynamics and protein–ligand dockings.
Original languageEnglish
Pages (from-to)1863-1868
JournalAngewandte Chemie - International Edition
Issue number7
Publication statusPublished - 12 Feb 2018


  • artificial metalloenzymes
  • cyclic imines
  • directed evolution
  • enzyme catalysis
  • transfer hydrogenation


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