Work

Development of a Universal Platform for the Heterologous Expression of Bidirectional [Ni–Fe]-Hydrogenases in E. coli

Year: 2025

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide Bidirectional [Ni–Fe]-hydrogenases are useful tools for integrating hydrogen into existing chemical processes by utilizing H 2 to regenerate expensive cofactors such as NAD(P)H. One enzyme broadly applied to this purpose is the soluble [Ni–Fe]-hydrogenase from Cupriavidus necator ( Cn SH). However, the homologous production of Cn SH suffers from slow growth rates and complex growth medium requirements of the native host. In the present study, we developed a simple approach for the production of Cn SH in Escherichia coli based on the coexpression of the maturation factors from C. necator . By optimizing the artificial operons coding for the hydrogenase proteins as well as the maturation factors, we were able to produce Cn SH with similar yields and activities compared to the native host. Additionally, we used our system to express three functional novel soluble [Ni–Fe]-hydrogenases, demonstrating its applicability for future enzyme screening and discovery.

Source: ACS Synthetic Biology

DOI: 10.1021/acssynbio.5c00150

Authors: Dominik L. Siebert, Frank Sargent, Ammar Al-Shameri, Volker Sieber

Institutions: Technical University of Munich, Newcastle University, The University of Queensland

Language: English

Topic: Metalloenzymes and iron-sulfur proteins

Subfield: Renewable Energy, Sustainability and the Environment

Field: Energy

Domain: Physical Sciences

Open Access status: hybrid

Estimated APC paid: $4,500

Funder: Bundesministerium für Bildung und Forschung