Hydrogenase-based oxidative biocatalysis without oxygen

Nature Communications, vol. 14, pp. 2693

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.

Authors 5

1. Ammar Al‐Shameri

   Technical University of Munich

   Affiliation as printed

   Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany

2. Dominik L. Siebert

   Technical University of Munich

   Affiliation as printed

   Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany

3. Samuel Sutiono

   Technical University of Munich

   Affiliation as printed

   Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany

4. Lars Lauterbach Aachen Institute of Applied Microbiology

   RWTH Aachen University

   Affiliation as printed

   RWTH Universität Aachen, Institute of Applied Microbiology, Worringerweg 1, 52074, Aachen, Germany

5. Volker Sieber

   The University of Queensland · Technical University of Munich

   Affiliation as printed

   Catalytic Research Center, Technical University of Munich, Ernst-Otto-Fischer-Straße 1, 85748, Garching, Germany. sieber@tum.de

   Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany. sieber@tum.de

   School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, QLD, 4072, Australia. sieber@tum.de

   SynBiofoundry@TUM, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany. sieber@tum.de

   Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315, Straubing, Germany