75424 20220621094623.0 doi 10.1039/C7CP05904C sideral 102550 ART-2020-102550 eng Carro, Juan Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase 2020 Access copy available to the general public Unrestricted 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. info:eu-repo/grantAgreement/EC/FP7/613549/EU/Optimized oxidoreductases for medium and large scale industrial biotransformations/INDOX info:eu-repo/grantAgreement/EC/H2020/720297/EU/New enzymatic oxidation/oxyfunctionalization technologies for added value bio-based products/EnzOx2 This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 720297-EnzOx2 info:eu-repo/grantAgreement/ES/MINECO/BIO2016-75183-P info:eu-repo/grantAgreement/ES/MINECO/NOESIS-BIO2014-56388-R info:eu-repo/semantics/openAccess All rights reserved http://www.europeana.eu/rights/rr-f/ 3.676 2020 PHYSICS, ATOMIC, MOLECULAR & CHEMICAL 8 / 37 = 0.216 2020 Q1 T1 CHEMISTRY, PHYSICAL 77 / 162 = 0.475 2020 Q2 T2 1.052 2020 Physics and Astronomy (miscellaneous) 2020 Q1 Physical and Theoretical Chemistry 2020 Q1 info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion Martínez-Julve, Marta Universidad de Zaragoza (orcid)0000-0001-9047-0046 Medina, Milagros Universidad de Zaragoza (orcid)0000-0001-8743-0182 Martínez, Angel T. Ferreira, Patricia Universidad de Zaragoza (orcid)0000-0003-4076-6118 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 19 (2020), 28666 Phys. chem. chem. phys. Physical Chemistry Chemical Physics 1463-9076 929175 http://zaguan.unizar.es/record/75424/files/texto_completo.pdf Postprint 111909 http://zaguan.unizar.es/record/75424/files/texto_completo.jpg?subformat=icon icon Postprint oai:zaguan.unizar.es:75424 articulos driver 2022-06-21-09:39:58 ARTICLE