Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization

The focus of the present work is to find the optimal conditions for the activation of agarose beads with divinyl sulfone (DVS). The reactivity of the vinyl sulfone groups in the support was checked by the support capacity to react with ethylamine; via elemental analysis. In addition, trypsin was use...

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Autores: Abellanas Pérez, Pedro, Andrades, Diandra de, Alcántara León, Andrés Rafael, Polizeli, Maria de Lourdes Teixeira de Moraes, Rocha Martín, Javier, Fernández Lafuente, Roberto
Tipo de recurso: artículo
Fecha de publicación:2024
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/111280
Acceso en línea:https://hdl.handle.net/20.500.14352/111280
Access Level:acceso abierto
Palabra clave:577.1
577.15
577.2
539.199
Multipoint covalent enzyme immobilization
Optimization of the activation conditions
Divinyl sulfone
Bioquímica (Biología)
Biología molecular (Biología)
2403 Bioquímica
2415 Biología Molecular
2302.09 Enzimología
2302.91 Química de Macromoléculas Biológicas
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oai_identifier_str oai:docta.ucm.es:20.500.14352/111280
network_acronym_str ES
network_name_str España
repository_id_str
spelling Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilizationAbellanas Pérez, PedroAndrades, Diandra deAlcántara León, Andrés RafaelPolizeli, Maria de Lourdes Teixeira de MoraesRocha Martín, JavierFernández Lafuente, Roberto577.1577.15577.2539.199Multipoint covalent enzyme immobilizationOptimization of the activation conditionsDivinyl sulfoneBioquímica (Biología)Biología molecular (Biología)2403 Bioquímica2415 Biología Molecular2302.09 Enzimología2302.91 Química de Macromoléculas BiológicasThe focus of the present work is to find the optimal conditions for the activation of agarose beads with divinyl sulfone (DVS). The reactivity of the vinyl sulfone groups in the support was checked by the support capacity to react with ethylamine; via elemental analysis. In addition, trypsin was used as a model enzyme to test the immobilization and stabilization capabilities of the different supports. The higher the pH, the more vinyl sulfone groups are incorporated into the support, but lower reactivity versus ethylamine is observed. Too long activation times led to similar results. A N/S ratio of 1 means that all vinyl sulfone groups were reactive, and it was always lower than tis figure. The N in the support was 50 % of the amount observed for glyoxyl supports activated with ethylenediamine, suggesting the VS polymerization may be a likely explanation for this result. The higher N/S ratio in the support (modified with ethylamine), the higher the obtained stabilization, very likely by the lower polymerization of the vinyl sulfone on the support. We propose 360 mM divinyl sulfone, at pH 11.5 and 2 h as optimal conditions to reach the highest enzyme stabilization by immobilization in this support.ElsevierUniversidad Complutense de Madrid20242024-01-0120242024-01-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/111280reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022-136535OB-I00MICIU info:eu-repo grantAgreementAgencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022-139209OB-C22open accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial 4.0 Internationalhttp://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/1112802026-06-02T12:44:21Z
dc.title.none.fl_str_mv Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
title Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
spellingShingle Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
Abellanas Pérez, Pedro
577.1
577.15
577.2
539.199
Multipoint covalent enzyme immobilization
Optimization of the activation conditions
Divinyl sulfone
Bioquímica (Biología)
Biología molecular (Biología)
2403 Bioquímica
2415 Biología Molecular
2302.09 Enzimología
2302.91 Química de Macromoléculas Biológicas
title_short Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
title_full Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
title_fullStr Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
title_full_unstemmed Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
title_sort Optimizing the activation of agarose beads with divinyl sulfone for enzyme immobilization and stabilization
dc.creator.none.fl_str_mv Abellanas Pérez, Pedro
Andrades, Diandra de
Alcántara León, Andrés Rafael
Polizeli, Maria de Lourdes Teixeira de Moraes
Rocha Martín, Javier
Fernández Lafuente, Roberto
author Abellanas Pérez, Pedro
author_facet Abellanas Pérez, Pedro
Andrades, Diandra de
Alcántara León, Andrés Rafael
Polizeli, Maria de Lourdes Teixeira de Moraes
Rocha Martín, Javier
Fernández Lafuente, Roberto
author_role author
author2 Andrades, Diandra de
Alcántara León, Andrés Rafael
Polizeli, Maria de Lourdes Teixeira de Moraes
Rocha Martín, Javier
Fernández Lafuente, Roberto
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 577.1
577.15
577.2
539.199
Multipoint covalent enzyme immobilization
Optimization of the activation conditions
Divinyl sulfone
Bioquímica (Biología)
Biología molecular (Biología)
2403 Bioquímica
2415 Biología Molecular
2302.09 Enzimología
2302.91 Química de Macromoléculas Biológicas
topic 577.1
577.15
577.2
539.199
Multipoint covalent enzyme immobilization
Optimization of the activation conditions
Divinyl sulfone
Bioquímica (Biología)
Biología molecular (Biología)
2403 Bioquímica
2415 Biología Molecular
2302.09 Enzimología
2302.91 Química de Macromoléculas Biológicas
description The focus of the present work is to find the optimal conditions for the activation of agarose beads with divinyl sulfone (DVS). The reactivity of the vinyl sulfone groups in the support was checked by the support capacity to react with ethylamine; via elemental analysis. In addition, trypsin was used as a model enzyme to test the immobilization and stabilization capabilities of the different supports. The higher the pH, the more vinyl sulfone groups are incorporated into the support, but lower reactivity versus ethylamine is observed. Too long activation times led to similar results. A N/S ratio of 1 means that all vinyl sulfone groups were reactive, and it was always lower than tis figure. The N in the support was 50 % of the amount observed for glyoxyl supports activated with ethylenediamine, suggesting the VS polymerization may be a likely explanation for this result. The higher N/S ratio in the support (modified with ethylamine), the higher the obtained stabilization, very likely by the lower polymerization of the vinyl sulfone on the support. We propose 360 mM divinyl sulfone, at pH 11.5 and 2 h as optimal conditions to reach the highest enzyme stabilization by immobilization in this support.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024-01-01
2024
2024-01-01
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/20.500.14352/111280
url https://hdl.handle.net/20.500.14352/111280
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022-136535OB-I00
MICIU info:eu-repo grantAgreement
Agencia Estatal de Investigación http://dx.doi.org/10.13039/501100011033 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022-139209OB-C22
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
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 4.0 International
http://creativecommons.org/licenses/by-nc/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,811543

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