Laccase-luminol chemiluminescence system: an investigation of substrate inhibition

Chemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behaviour has not been characterized. This study was conducted to characterize the laccase-luminol system...

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Autores: Sánchez Trasviña, Calef, Galindo Estrada, José Daniel, Tinoco Valencia, Raunel, Serrano Carreón, Leobardo, Rito Palomares, Marco, Willson, Richard C, Mayolo Deloisa, Karla P.
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/329772
Acceso en línea:http://hdl.handle.net/10261/329772
https://api.elsevier.com/content/abstract/scopus_id/85148459481
Access Level:acceso abierto
Palabra clave:Biosensor
Chemiluminescence
Laccase
Luminol
Substrate inhibition
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spelling Laccase-luminol chemiluminescence system: an investigation of substrate inhibitionSánchez Trasviña, CalefGalindo Estrada, José DanielTinoco Valencia, RaunelSerrano Carreón, LeobardoRito Palomares, MarcoWillson, Richard CMayolo Deloisa, Karla P.BiosensorChemiluminescenceLaccaseLuminolSubstrate inhibitionChemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behaviour has not been characterized. This study was conducted to characterize the laccase-luminol system, determine its kinetic parameters, and analyze the effects of protein and OH- concentration on the CL signal. Laccase from Coriolopsis gallica was combined with different concentrations of luminol (125 nM to 4 mM), and the enzyme kinetics were evaluated using diverse kinetic models. The laccase-luminol system was able to produce CL without an intermediate molecule, but it exhibited substrate-inhibition behaviour. A two-site random model was used and suggested that when the first luminol molecule was bound to the active site, laccase affinity for the second luminol molecule was increased. This inhibition effect could be avoided using a low luminol concentration. At 5 μM luminol concentration, 1 mg/ml (0.13 U) laccase is needed to achieve nearly 90% of the maximum CL signal, suggesting that the available luminol could not bind to all active sites. Furthermore, the concentration of NaOH negatively affected the CL signal. The laccase-luminol system represents an alternative to existing CL systems, with potential uses in molecular detection and quantification.Translational Omics and Bioengineering and Regenerative Medicine strategic focus groups of Tecnologico de Monterrey; CONACyT, Grant/Award Number: 492276With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000917-S).Peer reviewedWiley-VCHInstituto Tecnológico y de Estudios Superiores de MonterreyAgencia Estatal de Investigación (España)Consejo Nacional de Ciencia y Tecnología (México)Mayolo Deloisa, Karla P. [0000-0002-2826-2518]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202320232023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttp://hdl.handle.net/10261/329772https://api.elsevier.com/content/abstract/scopus_id/85148459481reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de investigación Científica y Técnica y de Innovación 2017-2020/CEX2019-000917-Shttp://doi.org/10.1002/bio.4460Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3297722026-05-22T06:33:51Z
dc.title.none.fl_str_mv Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
title Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
spellingShingle Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
Sánchez Trasviña, Calef
Biosensor
Chemiluminescence
Laccase
Luminol
Substrate inhibition
title_short Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
title_full Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
title_fullStr Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
title_full_unstemmed Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
title_sort Laccase-luminol chemiluminescence system: an investigation of substrate inhibition
dc.creator.none.fl_str_mv Sánchez Trasviña, Calef
Galindo Estrada, José Daniel
Tinoco Valencia, Raunel
Serrano Carreón, Leobardo
Rito Palomares, Marco
Willson, Richard C
Mayolo Deloisa, Karla P.
author Sánchez Trasviña, Calef
author_facet Sánchez Trasviña, Calef
Galindo Estrada, José Daniel
Tinoco Valencia, Raunel
Serrano Carreón, Leobardo
Rito Palomares, Marco
Willson, Richard C
Mayolo Deloisa, Karla P.
author_role author
author2 Galindo Estrada, José Daniel
Tinoco Valencia, Raunel
Serrano Carreón, Leobardo
Rito Palomares, Marco
Willson, Richard C
Mayolo Deloisa, Karla P.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto Tecnológico y de Estudios Superiores de Monterrey
Agencia Estatal de Investigación (España)
Consejo Nacional de Ciencia y Tecnología (México)
Mayolo Deloisa, Karla P. [0000-0002-2826-2518]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Biosensor
Chemiluminescence
Laccase
Luminol
Substrate inhibition
topic Biosensor
Chemiluminescence
Laccase
Luminol
Substrate inhibition
description Chemiluminescence (CL) reactions are widely used for the detection and quantification of many types of analytes. Laccase has previously been proposed in CL reactions; however, its light emission behaviour has not been characterized. This study was conducted to characterize the laccase-luminol system, determine its kinetic parameters, and analyze the effects of protein and OH- concentration on the CL signal. Laccase from Coriolopsis gallica was combined with different concentrations of luminol (125 nM to 4 mM), and the enzyme kinetics were evaluated using diverse kinetic models. The laccase-luminol system was able to produce CL without an intermediate molecule, but it exhibited substrate-inhibition behaviour. A two-site random model was used and suggested that when the first luminol molecule was bound to the active site, laccase affinity for the second luminol molecule was increased. This inhibition effect could be avoided using a low luminol concentration. At 5 μM luminol concentration, 1 mg/ml (0.13 U) laccase is needed to achieve nearly 90% of the maximum CL signal, suggesting that the available luminol could not bind to all active sites. Furthermore, the concentration of NaOH negatively affected the CL signal. The laccase-luminol system represents an alternative to existing CL systems, with potential uses in molecular detection and quantification.
publishDate 2023
dc.date.none.fl_str_mv 2023
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/329772
https://api.elsevier.com/content/abstract/scopus_id/85148459481
url http://hdl.handle.net/10261/329772
https://api.elsevier.com/content/abstract/scopus_id/85148459481
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de investigación Científica y Técnica y de Innovación 2017-2020/CEX2019-000917-S
http://doi.org/10.1002/bio.4460
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Wiley-VCH
publisher.none.fl_str_mv Wiley-VCH
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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