As a highly electrophilic moiety capable of oxidizing
a variety
of small organic molecules and biomolecules, flavin is an important
prosthetic group in many enzymes. Upon oxidation of the substrate,
flavin is converted into its reduced (dihydrogenated) form. The catalytic
cycle is completed through oxidation back to the oxidized form, thus
restoring the enzyme’s oxidizing capability. While it has been
firmly established that oxidation of the reduced form of flavin is
cast by molecular oxygen, yielding oxidized flavin and hydrogen peroxide,
the mechanism of this process is still poorly understood. Herein,
we investigate the radical mechanism, which is one of the possible
reaction mechanisms, by quantum chemical calculations. Because molecular
oxygen exists as a triplet in its electronic ground state, whereas
the products are singlets, the reaction is accompanied by hopping
between electronic surfaces. We find that the rate-limiting factor
of flavin oxidation is likely associated with the change in the spin
state of the system. By considering several possible reactions involving
flavin and its derivatives in the radical form and by examining the
corresponding parts of the potential energy surface in various spin
states, we estimate the effective barrier of the kinetically and thermodynamically
preferred variant of flavin oxidation to be about 15 kcal/mol in the
gas phase and about 7 kcal/mol in a polar (aqueous) environment.
This is in agreement with kinetic studies of the corresponding monoamine
oxidase enzymes, confirming the radical mechanism as a viable option
for flavin regeneration in enzymes.