Electronic and photolytic properties of hydridocobalamin

Hydridocobalamin (HCbl), is a known member of the B₁₂ family of molecules (cobalamins, Cbls) yet unlike other well-studied Cbls, little is known of the electronic and photolytic properties of this species. Interest in HCbl has increased significantly in recent years when at least three experimentally proposed mechanisms implicate HCbl as an intermediary in the photoreaction of coenzyme B₁₂-dependent photoreceptor CarH. Specifically, cleavage of the Co-C_5' bond of coenzyme B₁₂ could lead to a β-hydride or β‑hydrogen elimination reaction to form HCbl. HCbl is known to be a transient species where the oxidation state of the Co is variable; Co(I)-H⁺ ↔ Co(II)-H ↔ Co(III)-H^-. Further, HCbl is a very unstable with a pKa of ~1. This complicates experimental studies and to the best of our knowledge there are no available crystal structures of HCbl - either for the isolated molecule or bound to an enzyme. In this study, the electronic structure, photolytic properties, and reactivity of HCbl were explored to determine the preferred oxidation state as well as its potential role in the formation of the photoproduct in CarH. Natural bond orbital (NBO) analysis was performed to determine the oxidation state of Co in isolated HCbl. Based on the NBO analysis of HCbl, Co clearly had excess negative charge, which is in stark contrast to other alkylCbls where the Co ion is marked by significant positive charge. In sum, NBO results indicate that the CoH bond is strongly polarized and almost ionic. It can be described as protonated Co(I). In addition, DFT was used to explore the bond dissociation energy of HCbl based on homolytic cleavage of the CoH bond. TD-DFT calculations were used to compare computed electronic transitions to the experimentally determined absorption spectrum. The photoreaction of CarH was explored using an isolated model system and a pathway for hydrogen transfer was found. Finally, quantum mechanics/molecular mechanics (QM/MM) calculations were employed to investigate the formation of HCbl in CarH.