Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 57, Issue 7, Pages 1863-1868Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201711016
Keywords
artificial metalloenzymes; cyclic imines; directed evolution; enzyme catalysis; transfer hydrogenation
Categories
Funding
- Swiss Nanoscience Institute
- ERC grant (DrEAM)
- SNF [200020 162348]
- Spanish grant [CTQ2014-54071-P]
- Generalitat de Catalunya grant [2014SGR989]
- COST Action [CM1306]
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available