On how the binding cavity of AsqJ dioxygenase controls the desaturation reaction regioselectivity: a QM/MM study

ABSTRACT: The Fe(II)/2-oxoglutarate-dependent dioxygenase AsqJ from Aspergillus nidulans catalyses two pivotal steps in the synthesis of quinolone antibiotic 4′-methoxyviridicatin, i.e., desaturation and epoxidation of a benzodiazepinedione. The previous experimental results signal that, during the desaturation reaction, hydrogen atom transfer (HAT) from the benzylic carbon atom (C10) is a more likely step to initiate the reaction than the alternative HAT from the ring moiety (C3 atom). To unravel the origins of this regioselectivity and to explain why the observed reaction is desaturation and not the “default” hydroxylation, we performed a QM/MM study on the reaction catalysed by AsqJ. Herein, we report results that complement the experimental findings and suggest that HAT at the C10 position is the preferred reaction due to favourable interactions between the substrate and the binding cavity that compensate for the relatively high intrinsic barrier associated with the process. For the resultant radical intermediate, the desaturation/hydroxylation selectivity is governed by electronic properties of the reactants, i.e., the energy gap between the orbital that hosts the unpaired electron and the sigma orbital for the C–H bond as well as the gap between the orbitals mixing in transition state structures for each elementary step. GRAPHICAL ABSTRACT: Regiospecificity of the AsqJ dehydrogenation reaction is dictated by substrate–protein interactions. 82 × 44 mm (300 × 300 dpi) [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00775-018-1575-3) contains supplementary material, which is available to authorized users.