Please use this identifier to cite or link to this item: http://dx.doi.org/10.14279/depositonce-12023
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Main Title: Dihydrogen-driven NADPH recycling in imine reduction and P450-catalyzed oxidations mediated by an engineered O2-tolerant hydrogenase
Author(s): Preissler, Janina
Reeve, Holly A.
Zhu, Tianze
Nicholson, Jake
Urata, Kouji
Lauterbach, Lars
Wong, Luet L.
Vincent, Kylie A.
Lenz, Oliver
Type: Article
Language Code: en
Abstract: The O2-tolerant NAD+-reducing hydrogenase (SH) from Ralstonia eutropha (Cupriavidus necator) has already been applied in vitro and in vivo for H2-driven NADH recycling in coupled enzymatic reactions with various NADH-dependent oxidoreductases. To expand the scope for application in NADPH-dependent biocatalysis, we introduced changes in the NAD+-binding pocket of the enzyme by rational mutagenesis, and generated a variant with significantly higher affinity for NADP+ than for the natural substrate NAD+, while retaining native O2-tolerance. The applicability of the SH variant in H2-driven NADPH supply was demonstrated by the full conversion of 2-methyl-1-pyrroline into a single enantiomer of 2-methylpyrrolidine catalysed by a stereoselective imine reductase. In an even more challenging reaction, the SH supported a cytochrome P450 monooxygenase for the oxidation of octane under safe H2/O2 mixtures. Thus, the re-designed SH represents a versatile platform for atom-efficient, H2-driven cofactor recycling in biotransformations involving NADPH-dependent oxidoreductases.
URI: https://depositonce.tu-berlin.de/handle/11303/13228
http://dx.doi.org/10.14279/depositonce-12023
Issue Date: 16-Jun-2020
Date Available: 10-Jun-2021
DDC Class: 540 Chemie und zugeordnete Wissenschaften
Subject(s): hydrogenase
metalloenzyme
cytochrome P450
monooxygenase
oxidoreductase
imine reductase
octane oxidation
nicotinamide cofactor
NADH
cofactor recycling
Sponsor/Funder: DFG, 390540038, EXC 2008: Unifying Systems in Catalysis "UniSysCat"
TU Berlin, Open-Access-Mittel – 2020
License: https://creativecommons.org/licenses/by/4.0/
Journal Title: ChemCatChem
Publisher: Wiley
Publisher Place: New York, NY
Volume: 12
Issue: 19
Publisher DOI: 10.1002/cctc.202000763
Page Start: 4853
Page End: 4861
EISSN: 1867-3899
ISSN: 1867-3880
Appears in Collections:FG Physikalische Chemie / Biophysikalische Chemie » Publications

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