Rodríguez-Maciá Patricia, Reijerse Edward J, van Gastel Maurice, DeBeer Serena, Lubitz Wolfgang, Rüdiger Olaf, Birrell James A
Max Planck Institute for Chemical Energy Conversion , Stiftstraße 34-36 , D-45470 Mülheim an der Ruhr , Germany.
Max-Planck-Institut für Kohlenforschung , Kaiser-Wilhelm-Platz 1 , D-45470 Mülheim an der Ruhr , Germany.
J Am Chem Soc. 2018 Aug 1;140(30):9346-9350. doi: 10.1021/jacs.8b04339. Epub 2018 Jul 19.
[FeFe] hydrogenases catalyze proton reduction and hydrogen oxidation with high rates and efficiency under physiological conditions, but are highly oxygen sensitive. The [FeFe] hydrogenase from Desulfovibrio desulfuricans ( DdHydAB) can be purified under air in an oxygen stable inactive state H. The formation of the H state in vitro allows the handling of hydrogenases in air, making their implementation in biotechnological applications more feasible. Here, we report a simple and robust protocol for the formation of the H state in DdHydAB and the [FeFe] hydrogenase from Chlamydomonas reinhardtii, which is based on high potential inactivation in the presence of sulfide.
[铁铁]氢化酶在生理条件下能高效催化质子还原和氢氧化反应，但对氧气高度敏感。来自脱硫脱硫弧菌的[铁铁]氢化酶（DdHydAB）可在空气中纯化，处于氧稳定的无活性状态H。在体外形成H状态可使氢化酶在空气中进行处理，从而使其在生物技术应用中的实施更具可行性。在此，我们报告了一种简单且可靠的方案，用于在DdHydAB和莱茵衣藻的[铁铁]氢化酶中形成H状态，该方案基于在硫化物存在下的高电位失活。
