Projects per year
Abstract
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-IREDMs
variant and molecular dynamics (MD) simulations to rationalize
the inverted stereoselectivity of the quintuple variant.
Original language | English |
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Journal | ChemCatChem |
Early online date | 8 Oct 2021 |
DOIs | |
Publication status | E-pub ahead of print - 8 Oct 2021 |
Bibliographical note
© 2021 The Authors.Projects
- 1 Finished
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X-ray Diffraction Equipment for Macromolecular Crystallography at York
Davies, G. J. (Principal investigator), Antson, A. A. (Co-investigator), Brzozowski, A. M. (Co-investigator), Cowtan, K. (Co-investigator), Fascione, M. A. (Co-investigator), Grogan, G. J. (Co-investigator), Plevin, M. J. (Co-investigator), Thomas, G. H. (Co-investigator), Wilkinson, A. J. (Co-investigator), Willems, L. I. (Co-investigator) & Wilson, K. S. (Co-investigator)
BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL)
18/09/20 → 17/09/22
Project: Research project (funded) › Research