Siebert Dominik L, Sargent Frank, Al-Shameri Ammar, Sieber Volker
Chair of Chemistry of Biogenic Resources, TUM Campus Straubing for Biotechnology and Sustainability, Technical University of Munich, Schulgasse 16, 94315 Straubing, Germany.
Microbes in Health and Disease, Biosciences Institute, Newcastle University, Framlington Place, NE2 4HH Newcastle upon Tyne, U.K.
ACS Synth Biol. 2025 Jul 18;14(7):2710-2717. doi: 10.1021/acssynbio.5c00150. Epub 2025 Jun 16.
Bidirectional [Ni-Fe]-hydrogenases are useful tools for integrating hydrogen into existing chemical processes by utilizing H to regenerate expensive cofactors such as NAD(P)H. One enzyme broadly applied to this purpose is the soluble [Ni-Fe]-hydrogenase from (SH). However, the homologous production of 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 SH in based on the coexpression of the maturation factors from . By optimizing the artificial operons coding for the hydrogenase proteins as well as the maturation factors, we were able to produce 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.
双向[Ni-Fe]氢化酶是通过利用H来再生昂贵的辅因子（如NAD(P)H）从而将氢整合到现有化学过程中的有用工具。一种广泛应用于此目的的酶是来自嗜热栖热菌的可溶性[Ni-Fe]氢化酶（SH）。然而，SH的同源生产存在生长速率缓慢以及对天然宿主复杂生长培养基要求的问题。在本研究中，我们基于嗜热栖热菌成熟因子的共表达，开发了一种在大肠杆菌中生产SH的简单方法。通过优化编码氢化酶蛋白以及成熟因子的人工操纵子，我们能够以与天然宿主相似的产量和活性生产SH。此外，我们利用我们的系统表达了三种功能性新型可溶性[Ni-Fe]氢化酶，证明了其在未来酶筛选和发现中的适用性。
