Hydrogenase-based oxidative biocatalysis without oxygen

Ammar Al-Shameri, Dominik L. Siebert, Samuel Sutiono, Lars Lauterbach and Volker Sieber ()
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Ammar Al-Shameri: Technical University of Munich
Dominik L. Siebert: Technical University of Munich
Samuel Sutiono: Technical University of Munich
Lars Lauterbach: RWTH Universität Aachen, Institute of Applied Microbiology
Volker Sieber: Technical University of Munich

Nature Communications, 2023, vol. 14, issue 1, 1-8

Abstract: Abstract Biocatalysis-based synthesis can provide a sustainable and clean platform for producing chemicals. Many oxidative biocatalytic routes require the cofactor NAD+ as an electron acceptor. To date, NADH oxidase (NOX) remains the most widely applied system for NAD+ regeneration. However, its dependence on O2 implies various technical challenges in terms of O2 supply, solubility, and mass transfer. Here, we present the suitability of a NAD+ regeneration system in vitro based on H2 evolution. The efficiency of the hydrogenase-based system is demonstrated by integrating it into a multi-enzymatic cascade to produce ketoacids from sugars. The total NAD+ recycled using the hydrogenase system outperforms NOX in all different setups reaching up to 44,000 mol per mol enzyme. This system proves to be scalable and superior to NOX in terms of technical simplicity, flexibility, and total output. Furthermore, the system produces only green H2 as a by-product even in the presence of O2.

Date: 2023
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DOI: 10.1038/s41467-023-38227-9

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