Mechanistic principles of hydrogen evolution in the membrane-bound hydrogenase

Abstract

The membrane-bound hydrogenase (Mbh) from Pyrococcus furiosus is an archaeal member of the Complex I superfamily. It catalyzes the reduction of protons to H2 gas powered by a [NiFe] active site and transduces the free energy into proton pumping and Na+/H+-exchange across the membrane. Despite recent structural advances (1-4), the mechanistic principles of H2 catalysis and ion transport in Mbh remain elusive. Here we probe how the redox chemistry drives the proton reduction to H2 and how the catalysis couples to conformational dynamics in the membrane domain of Mbh. By combining large-scale quantum chemical density functional theory (DFT) and correlated ab initio wave function methods with atomistic molecular dynamics simulations, we show that the proton transfer reactions required for the catalysis are gated by electric field effects that direct the protons by water-mediated reactions from Glu21L towards the [NiFe] site, or alternatively along the nearby His75L pathway that also becomes energetically feasible in certain reaction steps. These local proton-coupled electron transfer (PCET) reactions induce conformational changes around the active site that provide a key coupling element via conserved loop structures to the ion transport activity. We find that H2 forms in a heterolytic proton reduction step, with spin crossovers tuning the energetics along key reaction steps. On a general level, our work showcases the role of electric fields in enzyme catalysis, and how these effects are employed by the [NiFe] active site of Mbh to drive the PCET reactions and ion transport. Significance statementHydrogen (H2) serves as a crucial solar fuel in renewable energy systems that can be efficiently produced by microbial hydrogenases. Here we probe the elusive mechanistic principles underlying the H2 production in the ancient membrane-bound hydrogenase (Mbh) from the thermophilic archaeon Pyrococcus furiosus. Distinct from other hydrogenases, Mbh not only produces H2, but it couples this activity with ion transport across a membrane that powers the archaeal energy metabolism. Our study elucidates key mechanistic principles underlying H2 production and shed light on energy transducing enzymes that led to the evolution of modern mitochondrial respiratory enzymes.