Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase

Aryl-alcohol oxidase (AAO) has demonstrated to be an enzyme with a bright future ahead due to its biotechnological potential in deracemisation of chiral compounds, production of bioplastic precursors and other reactions of interest. Expanding our understanding on the AAO reaction mechanisms, through...

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Authors: Carro, Juan, Amengual-Rigo, Pep, Sancho, Ferran, Medina, Milagros, Guallar, Víctor|||0000-0002-4580-1114, Ferreira, Patricia, Martinez, Angel T.
Format: article
Publication Date:2018
Country:España
Institution:Universitat Politècnica de Catalunya (UPC)
Repository:UPCommons. Portal del coneixement obert de la UPC
Language:English
OAI Identifier:oai:upcommons.upc.edu:2117/119386
Online Access:https://hdl.handle.net/2117/119386
https://dx.doi.org/10.1038/s41598-018-26445-x
Access Level:Open access
Keyword:Alcohol oxidoreductase
Enzyme activation
Aryl-alcohol oxidase (AAO)
Spectroscopic studies
Enzims
Àrees temàtiques de la UPC::Ciències de la salut
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spelling Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidaseCarro, JuanAmengual-Rigo, PepSancho, FerranMedina, MilagrosGuallar, Víctor|||0000-0002-4580-1114Ferreira, PatriciaMartinez, Angel T.Alcohol oxidoreductaseEnzyme activationAryl-alcohol oxidase (AAO)Spectroscopic studiesEnzimsÀrees temàtiques de la UPC::Ciències de la salutAryl-alcohol oxidase (AAO) has demonstrated to be an enzyme with a bright future ahead due to its biotechnological potential in deracemisation of chiral compounds, production of bioplastic precursors and other reactions of interest. Expanding our understanding on the AAO reaction mechanisms, through the investigation of its structure-function relationships, is crucial for its exploitation as an industrial biocatalyst. In this regard, previous computational studies suggested an active role for AAO Phe397 at the active-site entrance. This residue is located in a loop that partially covers the access to the cofactor forming a bottleneck together with two other aromatic residues. Kinetic and affinity spectroscopic studies, complemented with computational simulations using the recently developed adaptive-PELE technology, reveal that the Phe397 residue is important for product release and to help the substrates attain a catalytically relevant position within the active-site cavity. Moreover, removal of aromaticity at the 397 position impairs the oxygen-reduction activity of the enzyme. Experimental and computational findings agree very well in the timing of product release from AAO, and the simulations help to understand the experimental results. This highlights the potential of adaptive-PELE to provide answers to the questions raised by the empirical results in the study of enzyme mechanisms.This work was supported by the EnzOx2 project (H2020-BBI-PPP-2015-720297) of the European Joint Undertaking of Bio-based Industries (http://bbi-europe.eu), the INDOX project (KBBE-2013-7-613549) of the European Seventh Framework Programme, and the NOESIS (BIO2014-56388-R), vMutate (CTQ2016-79138-R) and FLADIMOTEC (BIO2016-75183-P) projects of the Spanish Ministry of Economy and Competitiveness. J.C. acknowledges a FPU fellowship (FPU2012-2041) from the Spanish Ministry of Education, Culture and Sports.Peer ReviewedNature Publishing Group20182018-05-2520182018-07-16journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/119386https://dx.doi.org/10.1038/s41598-018-26445-x29802285reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)InglésengEuropean Commission http://doi.org/10.13039/100010661 Horizon 2020 Framework Programme 720297 New enzymatic oxidation%2Foxyfunctionalization technologies for added value bio-based productsEuropean Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 613549 Optimized oxidoreductases for medium and large scale industrial biotransformationsMinisterio de Economía y Competitividad http://doi.org/10.13039/501100003329 BIO2014-56388-R NUEVAS ENZIMAS OXIDATIVAS PARA UNA INDUSTRIA SOSTENIBLEopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivs 3.0 Spainhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/1193862026-05-27T15:37:01Z
dc.title.none.fl_str_mv Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
title Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
spellingShingle Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
Carro, Juan
Alcohol oxidoreductase
Enzyme activation
Aryl-alcohol oxidase (AAO)
Spectroscopic studies
Enzims
Àrees temàtiques de la UPC::Ciències de la salut
title_short Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
title_full Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
title_fullStr Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
title_full_unstemmed Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
title_sort Multiple implications of an active site phenylalanine in the catalysis of aryl-alcohol oxidase
dc.creator.none.fl_str_mv Carro, Juan
Amengual-Rigo, Pep
Sancho, Ferran
Medina, Milagros
Guallar, Víctor|||0000-0002-4580-1114
Ferreira, Patricia
Martinez, Angel T.
author Carro, Juan
author_facet Carro, Juan
Amengual-Rigo, Pep
Sancho, Ferran
Medina, Milagros
Guallar, Víctor|||0000-0002-4580-1114
Ferreira, Patricia
Martinez, Angel T.
author_role author
author2 Amengual-Rigo, Pep
Sancho, Ferran
Medina, Milagros
Guallar, Víctor|||0000-0002-4580-1114
Ferreira, Patricia
Martinez, Angel T.
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Alcohol oxidoreductase
Enzyme activation
Aryl-alcohol oxidase (AAO)
Spectroscopic studies
Enzims
Àrees temàtiques de la UPC::Ciències de la salut
topic Alcohol oxidoreductase
Enzyme activation
Aryl-alcohol oxidase (AAO)
Spectroscopic studies
Enzims
Àrees temàtiques de la UPC::Ciències de la salut
description Aryl-alcohol oxidase (AAO) has demonstrated to be an enzyme with a bright future ahead due to its biotechnological potential in deracemisation of chiral compounds, production of bioplastic precursors and other reactions of interest. Expanding our understanding on the AAO reaction mechanisms, through the investigation of its structure-function relationships, is crucial for its exploitation as an industrial biocatalyst. In this regard, previous computational studies suggested an active role for AAO Phe397 at the active-site entrance. This residue is located in a loop that partially covers the access to the cofactor forming a bottleneck together with two other aromatic residues. Kinetic and affinity spectroscopic studies, complemented with computational simulations using the recently developed adaptive-PELE technology, reveal that the Phe397 residue is important for product release and to help the substrates attain a catalytically relevant position within the active-site cavity. Moreover, removal of aromaticity at the 397 position impairs the oxygen-reduction activity of the enzyme. Experimental and computational findings agree very well in the timing of product release from AAO, and the simulations help to understand the experimental results. This highlights the potential of adaptive-PELE to provide answers to the questions raised by the empirical results in the study of enzyme mechanisms.
publishDate 2018
dc.date.none.fl_str_mv 2018
2018-05-25
2018
2018-07-16
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2117/119386
https://dx.doi.org/10.1038/s41598-018-26445-x
29802285
url https://hdl.handle.net/2117/119386
https://dx.doi.org/10.1038/s41598-018-26445-x
identifier_str_mv 29802285
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv European Commission http://doi.org/10.13039/100010661 Horizon 2020 Framework Programme 720297 New enzymatic oxidation%2Foxyfunctionalization technologies for added value bio-based products
European Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 613549 Optimized oxidoreductases for medium and large scale industrial biotransformations
Ministerio de Economía y Competitividad http://doi.org/10.13039/501100003329 BIO2014-56388-R NUEVAS ENZIMAS OXIDATIVAS PARA UNA INDUSTRIA SOSTENIBLE
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivs 3.0 Spain
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivs 3.0 Spain
http://creativecommons.org/licenses/by-nc-nd/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:UPCommons. Portal del coneixement obert de la UPC
instname:Universitat Politècnica de Catalunya (UPC)
instname_str Universitat Politècnica de Catalunya (UPC)
reponame_str UPCommons. Portal del coneixement obert de la UPC
collection UPCommons. Portal del coneixement obert de la UPC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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