Calculating the chemical mechanism of nitrogenase: new working hypotheses

The enzyme nitrogenase converts N₂ to NH₃ with stoichiometry N₂ + 8H⁺ + 8e^- → 2NH₃ + H₂. The mechanism is chemically complex with multiple steps that must be consistent with much accumulated experimental information, including exchange of H₂ and N₂ and the N₂-dependent hydrogenation of D₂ to HD. Previous investigations have developed a collection of working hypotheses that guide ongoing density functional investigations of mechanistic steps and sequences. These include (i) hypotheses about the serial provision of protons and their conversion to H atoms bonded to S and Fe atoms of the FeMo-co catalytic site, (ii) the migration of H atoms over the surface of FeMo-co, (iii) the roles of His195, (iv) identification of three protein channels, one for the ingress of N₂, a separate pathway for the passage of exogenous H₂ (D₂) and product H₂ (HD), and a hydrophilic pathway for egress of product NH₃. Two additional working hypotheses are described in this paper. N₂ passing along the N₂ channel approaches and binds end-on to the exo coordination position of Fe2, with favourable energetics when FeMo-co is pre-hydrogenated. This exo-Fe2-N₂ is apparently not reduced but has a promotional role by expanding the reaction zone. A second N₂ can enter via the N₂ ingress channel and bind at the endo-Fe6 position, where it is surrounded by H atom donors suitable for the N₂ → NH₃ conversion. It is proposed that this endo-Fe6 position is also the binding site for H₂ (generated or exogenous), accounting for the competitive inhibition of N₂ reduction by H₂. The HD reaction occurs at the endo-Fe6 site, promoted by N₂ at the exo-Fe2 site. The second hypothesis concerns the most stable electronic states of FeMo-co with ligands bound at Fe2 and Fe6, and provides a protocol for management of electronic states in mechanism calculations.