Department of Chemistry-, Uppsala University, Lägerhyddsvägen 1, 75120, Uppsala, Sweden.
Angew Chem Int Ed Engl. 2018 Mar 1;57(10):2596-2599. doi: 10.1002/anie.201710740. Epub 2018 Feb 6.
EPR spectroscopy reveals the formation of two different semi-synthetic hydrogenases in vivo. [FeFe] hydrogenases are metalloenzymes that catalyze the interconversion of molecular hydrogen and protons. The reaction is catalyzed by the H-cluster, consisting of a canonical iron-sulfur cluster and an organometallic [2Fe] subsite. It was recently shown that the enzyme can be reconstituted with synthetic cofactors mimicking the composition of the [2Fe] subsite, resulting in semi-synthetic hydrogenases. Herein, we employ EPR spectroscopy to monitor the formation of two such semi-synthetic enzymes in whole cells. The study provides the first spectroscopic characterization of semi-synthetic hydrogenases in vivo, and the observation of two different oxidized states of the H-cluster under intracellular conditions. Moreover, these findings underscore how synthetic chemistry can be a powerful tool for manipulation and examination of the hydrogenase enzyme under in vivo conditions.
EPR 光谱揭示了两种不同的半合成氢化酶在体内的形成。[FeFe]氢化酶是一种金属酶，能够催化氢气和质子的相互转化。该反应由 H 簇催化，H 簇由一个典型的铁-硫簇和一个有机金属[2Fe]亚位点组成。最近的研究表明，该酶可以用模拟[2Fe]亚位点组成的合成辅因子进行重组，从而产生半合成氢化酶。在此，我们采用 EPR 光谱法在全细胞中监测两种此类半合成酶的形成。该研究首次对体内半合成氢化酶进行了光谱表征，并观察到在细胞内条件下 H 簇的两种不同氧化态。此外，这些发现强调了合成化学如何成为在体内条件下操纵和研究氢化酶的有力工具。
