Kinetic and Spectroscopic Studies of Bicupin Oxalate Oxidase and Putative Active Site Mutants
The detailed manganese-dependent chemistry employed by oxalate decarboxylase (OxDC) to catalyze the nonoxidative decarboxylation of oxalic acid remains poorly understood. For example, enzyme activity requires the presence of dioxygen even though this compound is not a formal substrate in the reaction. We now report density functional theory (DFT) calculations upon a series of hypothetical OxDC active site model structures. Our results suggest that the function of the metal ion may be to position dioxygen and oxalate such that electrons can be shuttled directly between these species, thereby removing the need for the existence of Mn(III) as an intermediate in the mechanism. These calculations also indicate that the intrinsic, gas-phase reactivity of the Bacillus subtilis oxalate decarboxylase active center is to oxidize oxalate. Since this reactivity is not observed for OxDC, our DFT results suggest that protein environment modulates the intrinsic metallocenter reactivity, presumably by affecting the electronic distribution at the manganese center during catalysis.