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Engineering a Formate Dehydrogenase for NADPH Regeneration.

Authors :
Ma W
Geng Q
Chen C
Zheng YC
Yu HL
Xu JH
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