Surface coverage dictates the surface bio-activity of d-amino acid oxidase

Autores
Herrera, Elisa Gabriela; Giacomelli, Carla Eugenia
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
This work presents a systematic study on the relationship between the adsorption mechanism and the surface bio-activity of d-amino acid oxidase (pkDAAO). This rational approach is based on measuring the characteristic filling and relaxation times under different experimental conditions. With such a goal, real-time adsorption–desorption experiments at different degrees of surface coverage were performed tuning the electrostatic and hydrophobic interactions by changing the pH condition for the adsorption and the substrate properties (silica or gold). Surface bio-activity was measured in situ by amperometry using the bio-functional surface as the working electrode and ex situ by spectrophotometry. On both solid substrates, pkDAAO adsorption is a transport-controlled process, even under unfavorable electrostatic interactions (charged protein and substrate with the same sign) due to the high percentage of basic amino acids in the enzyme. On silica, the relaxation step is electrostatic in nature and occurs in the same time-scale as filling the surface when the substrate and the enzyme are oppositely charged at low surface coverage. Under unfavorable electrostatic conditions, the relaxation (if any) occurs at long time. Accordingly, the bio-activity of the native pkDAAO is preserved at any surface coverage. On gold, this step is driven by hydrophobic interactions (pH-independent) and the surface bio-activity is highly dependent on the degree of surface coverage. Under these conditions, the surface bio-activity is preserved only at high surfaces coverage. Our results clearly indicate that pkDAAO bio-functionalized surfaces cannot be coupled to amperometry because the analyte interferes the electrochemical signal. However, this simple bio-functionalized strategy can be joined to other detection methods.
Fil: Herrera, Elisa Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
Fil: Giacomelli, Carla Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
Materia
Silica
Gold
Daao
Adsorption
Filling Time
Surface Relaxation
Surface Denaturing
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/31356
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/31356
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Surface coverage dictates the surface bio-activity of d-amino acid oxidaseHerrera, Elisa GabrielaGiacomelli, Carla EugeniaSilicaGoldDaaoAdsorptionFilling TimeSurface RelaxationSurface Denaturinghttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1This work presents a systematic study on the relationship between the adsorption mechanism and the surface bio-activity of d-amino acid oxidase (pkDAAO). This rational approach is based on measuring the characteristic filling and relaxation times under different experimental conditions. With such a goal, real-time adsorption–desorption experiments at different degrees of surface coverage were performed tuning the electrostatic and hydrophobic interactions by changing the pH condition for the adsorption and the substrate properties (silica or gold). Surface bio-activity was measured in situ by amperometry using the bio-functional surface as the working electrode and ex situ by spectrophotometry. On both solid substrates, pkDAAO adsorption is a transport-controlled process, even under unfavorable electrostatic interactions (charged protein and substrate with the same sign) due to the high percentage of basic amino acids in the enzyme. On silica, the relaxation step is electrostatic in nature and occurs in the same time-scale as filling the surface when the substrate and the enzyme are oppositely charged at low surface coverage. Under unfavorable electrostatic conditions, the relaxation (if any) occurs at long time. Accordingly, the bio-activity of the native pkDAAO is preserved at any surface coverage. On gold, this step is driven by hydrophobic interactions (pH-independent) and the surface bio-activity is highly dependent on the degree of surface coverage. Under these conditions, the surface bio-activity is preserved only at high surfaces coverage. Our results clearly indicate that pkDAAO bio-functionalized surfaces cannot be coupled to amperometry because the analyte interferes the electrochemical signal. However, this simple bio-functionalized strategy can be joined to other detection methods.Fil: Herrera, Elisa Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; ArgentinaFil: Giacomelli, Carla Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; ArgentinaElsevier Science2014-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/31356Giacomelli, Carla Eugenia; Herrera, Elisa Gabriela; Surface coverage dictates the surface bio-activity of d-amino acid oxidase; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 117; 3-2014; 296-3020927-7765CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2014.02.050info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0927776514001179info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:51:04Zoai:ri.conicet.gov.ar:11336/31356instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:51:05.23CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Surface coverage dictates the surface bio-activity of d-amino acid oxidase
title Surface coverage dictates the surface bio-activity of d-amino acid oxidase
spellingShingle Surface coverage dictates the surface bio-activity of d-amino acid oxidase
Herrera, Elisa Gabriela
Silica
Gold
Daao
Adsorption
Filling Time
Surface Relaxation
Surface Denaturing
title_short Surface coverage dictates the surface bio-activity of d-amino acid oxidase
title_full Surface coverage dictates the surface bio-activity of d-amino acid oxidase
title_fullStr Surface coverage dictates the surface bio-activity of d-amino acid oxidase
title_full_unstemmed Surface coverage dictates the surface bio-activity of d-amino acid oxidase
title_sort Surface coverage dictates the surface bio-activity of d-amino acid oxidase
dc.creator.none.fl_str_mv Herrera, Elisa Gabriela
Giacomelli, Carla Eugenia
author Herrera, Elisa Gabriela
author_facet Herrera, Elisa Gabriela
Giacomelli, Carla Eugenia
author_role author
author2 Giacomelli, Carla Eugenia
author2_role author
dc.subject.none.fl_str_mv Silica
Gold
Daao
Adsorption
Filling Time
Surface Relaxation
Surface Denaturing
topic Silica
Gold
Daao
Adsorption
Filling Time
Surface Relaxation
Surface Denaturing
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv This work presents a systematic study on the relationship between the adsorption mechanism and the surface bio-activity of d-amino acid oxidase (pkDAAO). This rational approach is based on measuring the characteristic filling and relaxation times under different experimental conditions. With such a goal, real-time adsorption–desorption experiments at different degrees of surface coverage were performed tuning the electrostatic and hydrophobic interactions by changing the pH condition for the adsorption and the substrate properties (silica or gold). Surface bio-activity was measured in situ by amperometry using the bio-functional surface as the working electrode and ex situ by spectrophotometry. On both solid substrates, pkDAAO adsorption is a transport-controlled process, even under unfavorable electrostatic interactions (charged protein and substrate with the same sign) due to the high percentage of basic amino acids in the enzyme. On silica, the relaxation step is electrostatic in nature and occurs in the same time-scale as filling the surface when the substrate and the enzyme are oppositely charged at low surface coverage. Under unfavorable electrostatic conditions, the relaxation (if any) occurs at long time. Accordingly, the bio-activity of the native pkDAAO is preserved at any surface coverage. On gold, this step is driven by hydrophobic interactions (pH-independent) and the surface bio-activity is highly dependent on the degree of surface coverage. Under these conditions, the surface bio-activity is preserved only at high surfaces coverage. Our results clearly indicate that pkDAAO bio-functionalized surfaces cannot be coupled to amperometry because the analyte interferes the electrochemical signal. However, this simple bio-functionalized strategy can be joined to other detection methods.
Fil: Herrera, Elisa Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
Fil: Giacomelli, Carla Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Físico-química de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Investigaciones en Físico-química de Córdoba; Argentina
description This work presents a systematic study on the relationship between the adsorption mechanism and the surface bio-activity of d-amino acid oxidase (pkDAAO). This rational approach is based on measuring the characteristic filling and relaxation times under different experimental conditions. With such a goal, real-time adsorption–desorption experiments at different degrees of surface coverage were performed tuning the electrostatic and hydrophobic interactions by changing the pH condition for the adsorption and the substrate properties (silica or gold). Surface bio-activity was measured in situ by amperometry using the bio-functional surface as the working electrode and ex situ by spectrophotometry. On both solid substrates, pkDAAO adsorption is a transport-controlled process, even under unfavorable electrostatic interactions (charged protein and substrate with the same sign) due to the high percentage of basic amino acids in the enzyme. On silica, the relaxation step is electrostatic in nature and occurs in the same time-scale as filling the surface when the substrate and the enzyme are oppositely charged at low surface coverage. Under unfavorable electrostatic conditions, the relaxation (if any) occurs at long time. Accordingly, the bio-activity of the native pkDAAO is preserved at any surface coverage. On gold, this step is driven by hydrophobic interactions (pH-independent) and the surface bio-activity is highly dependent on the degree of surface coverage. Under these conditions, the surface bio-activity is preserved only at high surfaces coverage. Our results clearly indicate that pkDAAO bio-functionalized surfaces cannot be coupled to amperometry because the analyte interferes the electrochemical signal. However, this simple bio-functionalized strategy can be joined to other detection methods.
publishDate 2014
dc.date.none.fl_str_mv 2014-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/31356
Giacomelli, Carla Eugenia; Herrera, Elisa Gabriela; Surface coverage dictates the surface bio-activity of d-amino acid oxidase; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 117; 3-2014; 296-302
0927-7765
CONICET Digital
CONICET
url http://hdl.handle.net/11336/31356
identifier_str_mv Giacomelli, Carla Eugenia; Herrera, Elisa Gabriela; Surface coverage dictates the surface bio-activity of d-amino acid oxidase; Elsevier Science; Colloids and Surfaces B: Biointerfaces; 117; 3-2014; 296-302
0927-7765
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.colsurfb.2014.02.050
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0927776514001179
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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