Evidence for C-H cleavage by an iron-superoxide complex in the glycol cleavage reaction catalyzed by myo-inositol oxygenase

myo-Inositol oxygenase (MIOX) activates O₂ at a mixed-valent nonheme diiron(II/III) cluster to effect oxidation of its cyclohexan(1,2,3,4,5, 6-hexa)-ol substrate [myo-inositol(MI)] by four electrons to D-glucuronate. Abstraction of hydrogen from C₁ by a formally (superoxo)diiron(III/ III) intermediate was previously proposed. Use of deuterium-labeled substrate, 1,2,3,4,5,6-[₂H]₆-MI (D₆-MI), has now permitted initial characterization of the C-H-cleaving intermediate. The MIOX·1,2,3,4,5,6-[²H]₆-MI complex reacts rapidly and reversibly with O₂ to form an intermediate, G, with a g = (2.05, 1.98, 1.90) EPR signal. The rhombic g-tensor and observed hyperfine coupling to ⁵⁷Fe are rationalized in terms of a (superoxo)diiron(III/III) structure with coordination of the superoxide to a single iron. G decays to H, the intermediate previously detected in the reaction with unlabeled substrate. This step is associated with a kinetic isotope effect of ≥5, showing that the superoxide-level complex does indeed cleave a C-H(D) bond of MI.