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Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase

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Abstract

The temperature dependence of hydride transfer from the substrate to the N5 of the FAD cofactor during the reductive half-reaction of Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both the pre-steady state reduction of the enzyme and its steady-state kinetics, with differently deuterated substrates, suggest an environmentally-coupled quantum-mechanical tunnelling process. Moreover, those kinetic data, along with the crystallographic structure of the enzyme in complex with a substrate analogue, indicate that AAO shows a pre-organized active site that would only require the approaching of the hydride donor and acceptor for the tunnelled transfer to take place. Modification of the enzyme's active-site architecture by replacement of Tyr92, a residue establishing hydrophobic interactions with the substrate analogue in the crystal structure, in the Y92F, Y92L and Y92W variants resulted in different temperature dependence patterns that indicated a role of this residue in modulating the transfer reaction.

Graphical abstract: Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase

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Publication details

The article was received on 29 Aug 2017, accepted on 09 Oct 2017 and first published on 11 Oct 2017


Article type: Paper
DOI: 10.1039/C7CP05904C
Citation: Phys. Chem. Chem. Phys., 2017, Advance Article
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    Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase

    J. Carro, M. Martínez-Júlvez, M. Medina, A. T. Martínez and P. Ferreira, Phys. Chem. Chem. Phys., 2017, Advance Article , DOI: 10.1039/C7CP05904C

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