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Engineering a Formate Dehydrogenase for NADPH Regeneration.
- Source :
-
Chembiochem : a European journal of chemical biology [Chembiochem] 2023 Oct 17; Vol. 24 (20), pp. e202300390. Date of Electronic Publication: 2023 Aug 28. - Publication Year :
- 2023
-
Abstract
- Nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH) constitute major hydrogen donors for oxidative/reductive bio-transformations. NAD(P)H regeneration systems coupled with formate dehydrogenases (FDHs) represent a dreamful method. However, most of the native FDHs are NAD <superscript>+</superscript> -dependent and suffer from insufficient reactivity compared to other enzymatic tools, such as glucose dehydrogenase. An efficient and competitive NADP <superscript>+</superscript> -utilizing FDH necessitates the availability and robustness of NADPH regeneration systems. Herein, we report the engineering of a new FDH from Candida dubliniensis (CdFDH), which showed no strict NAD <superscript>+</superscript> preference by a structure-guided rational/semi-rational design. A combinatorial mutant CdFDH-M4 (D197Q/Y198R/Q199N/A372S/K371T/▵Q375/K167R/H16L/K159R) exhibited 75-fold intensification of catalytic efficiency (k <subscript>cat</subscript> /K <subscript>m</subscript> ). Moreover, CdFDH-M4 has been successfully employed in diverse asymmetric oxidative/reductive processes with cofactor total turnover numbers (TTNs) ranging from 135 to 986, making it potentially useful for NADPH-required biocatalytic transformations.<br /> (© 2023 Wiley-VCH GmbH.)
Details
- Language :
- English
- ISSN :
- 1439-7633
- Volume :
- 24
- Issue :
- 20
- Database :
- MEDLINE
- Journal :
- Chembiochem : a European journal of chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 37455264
- Full Text :
- https://doi.org/10.1002/cbic.202300390