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http://dx.doi.org/10.18419/opus-14858
Autor(en): | Stockinger, Peter Borlinghaus, Niels Sharma, Mahima Aberle, Benjamin Grogan, Gideon Pleiss, Jürgen Nestl, Bettina M. |
Titel: | Inverting the stereoselectivity of an NADH‐dependent imine‐reductase variant |
Erscheinungsdatum: | 2021 |
Dokumentart: | Zeitschriftenartikel |
Seiten: | 5210-5215 |
Erschienen in: | ChemCatChem 13 (2021), S. 5210-5215 |
URI: | http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-148779 http://elib.uni-stuttgart.de/handle/11682/14877 http://dx.doi.org/10.18419/opus-14858 |
ISSN: | 1867-3899 1867-3880 |
Zusammenfassung: | Imine reductases (IREDs) offer biocatalytic routes to chiral amines and have a natural preference for the NADPH cofactor. In previous work, we reported enzyme engineering of the (R)‐selective IRED from Myxococcus stipitatus (NADH‐IRED‐Ms) yielding a NADH‐dependent variant with high catalytic efficiency. However, no IRED with NADH specificity and (S)‐selectivity in asymmetric reductions has yet been reported. Herein, we applied semi‐rational enzyme engineering to switch the selectivity of NADH‐IRED‐Ms. The quintuple variant A241V/H242Y/N243D/V244Y/A245L showed reverse stereopreference in the reduction of the cyclic imine 2‐methylpyrroline compared to the wild‐type and afforded the (S)‐amine product with >99 % conversion and 91 % enantiomeric excess. We also report the crystal‐structures of the NADPH‐dependent (R)‐IRED‐Ms wild‐type enzyme and the NADH‐dependent NADH‐IRED‐Ms variant and molecular dynamics (MD) simulations to rationalize the inverted stereoselectivity of the quintuple variant. |
Enthalten in den Sammlungen: | 03 Fakultät Chemie |
Dateien zu dieser Ressource:
Datei | Beschreibung | Größe | Format | |
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CCTC_CCTC202101057.pdf | 1,49 MB | Adobe PDF | Öffnen/Anzeigen |
Diese Ressource wurde unter folgender Copyright-Bestimmung veröffentlicht: Lizenz von Creative Commons