Solomon Joseph B, Lee Chi Chung, Liu Yiling A, Duffin Calder, Ribbe Markus W, Hu Yilin
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900.
Department of Chemistry, University of California, Irvine, CA 92697-2025.
Nat Catal. 2024 Oct;7(10):1130-1141. doi: 10.1038/s41929-024-01229-x. Epub 2024 Sep 19.
Heterologous expression of nitrogenase has been actively pursued because of the far-reaching impact of this enzyme on agriculture, energy and environment. Yet, isolation of an active two-component, metallocentre-containing nitrogenase from a non-diazotrophic host has yet to be accomplished. Here, we report the heterologous synthesis of an active Mo-nitrogenase by combining genes from and in . Metal, activity and EPR analyses demonstrate the integrity of the metallocentres in the purified nitrogenase enzyme; whereas growth, nanoSIMS and NMR experiments illustrate diazotrophic growth and N enrichment by the expression strain, as well as accumulation of extracellular ammonia upon deletion of the ammonia transporter that permits incorporation of thus-generated N into the cellular mass of a non-diazotrophic strain. As such, this study provides a crucial prototype system that could be optimized/modified to enable future transgenic expression and biotechnological adaptations of nitrogenase.
由于固氮酶对农业、能源和环境具有深远影响，人们一直在积极探索其异源表达。然而，从非固氮宿主中分离出具有活性的双组分、含金属中心的固氮酶尚未实现。在此，我们报告了通过将来自[具体来源1]和[具体来源2]的基因组合在[具体宿主]中实现活性钼固氮酶的异源合成。金属、活性和电子顺磁共振分析证明了纯化的固氮酶中金属中心的完整性；而生长、纳米二次离子质谱和核磁共振实验表明，[表达菌株名称]表达菌株具有固氮生长和氮富集能力，并且在缺失氨转运体后细胞外氨会积累，这使得由此产生的氮能够掺入非固氮[菌株名称]菌株的细胞物质中。因此，本研究提供了一个关键的原型系统，可对其进行优化/改进，以实现未来固氮酶的转基因表达和生物技术应用。
