Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization

Ficin extract has been immobilized using different supports: glyoxyl and Aspartic/1,6 hexamethylenediamine (Asp/HA) agarose beads. The latter was later submitted to glutaraldehyde modification to get covalent immobilization. The activities of these 3 kinds of biocatalysts were compared utilizing 4 d...

Descripción completa

Detalles Bibliográficos
Autores: González Vásquez, Alex D., Hocine, El Siar, Urzúa, Marcela, 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:20.500.14352/112066
Acceso en línea:https://hdl.handle.net/20.500.14352/112066
Access Level:acceso abierto
Palabra clave:577.1
577.15
547.94
544.478
Enzyme specificity
Conformational changes induced by immobilization
Enzyme tuning by immobilization
Bioquímica (Biología)
2403 Bioquímica
2302.09 Enzimología
2302.27 Proteínas
2210.01 Catálisis
id ES_be29f6a2e99a47f221a4f91e6b609bcd
oai_identifier_str oai:docta.ucm.es:20.500.14352/112066
network_acronym_str ES
network_name_str España
repository_id_str
spelling Changes in ficin specificity by different substrate proteins promoted by enzyme immobilizationGonzález Vásquez, Alex D.Hocine, El SiarUrzúa, MarcelaRocha Martín, JavierFernández Lafuente, Roberto577.1577.15547.94544.478Enzyme specificityConformational changes induced by immobilizationEnzyme tuning by immobilizationBioquímica (Biología)2403 Bioquímica2302.09 Enzimología2302.27 Proteínas2210.01 CatálisisFicin extract has been immobilized using different supports: glyoxyl and Aspartic/1,6 hexamethylenediamine (Asp/HA) agarose beads. The latter was later submitted to glutaraldehyde modification to get covalent immobilization. The activities of these 3 kinds of biocatalysts were compared utilizing 4 different substrates, casein, hemoglobin and bovine serum albumin and benzoyl-arginine-p-nitroanilide at pH 7 and 5. Using glyoxyl-agarose, the effect of enzyme-support reaction time on the activity versus the four substrates at both pH values was studied. Reaction time has been shown to distort the enzyme due to an increase in the number of covalent support-enzyme bonds. Surprisingly, for all the substrates and conditions the prolongation of the enzyme-support reaction did not imply a decrease in enzyme activity. Using the Asp/HA supports (with different amount of HA) differences in the effect on enzyme activity versus the different substrates are much more significant, while with some substrates the immobilization produced a decrease in enzyme activity, with in other cases the activity increased. These different effects are even increased after glutaraldehyde treatment. That way, the conformational changes induced by the biocatalyst immobilization or the chemical modification fully altered the enzyme protein specificity. This may also have some implications when following enzyme inactivation.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/20.500.14352/112066reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengMinisterio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022–136535OB-I00open 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:20.500.14352/1120662026-06-02T12:44:21Z
dc.title.none.fl_str_mv Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
title Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
spellingShingle Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
González Vásquez, Alex D.
577.1
577.15
547.94
544.478
Enzyme specificity
Conformational changes induced by immobilization
Enzyme tuning by immobilization
Bioquímica (Biología)
2403 Bioquímica
2302.09 Enzimología
2302.27 Proteínas
2210.01 Catálisis
title_short Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
title_full Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
title_fullStr Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
title_full_unstemmed Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
title_sort Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization
dc.creator.none.fl_str_mv González Vásquez, Alex D.
Hocine, El Siar
Urzúa, Marcela
Rocha Martín, Javier
Fernández Lafuente, Roberto
author González Vásquez, Alex D.
author_facet González Vásquez, Alex D.
Hocine, El Siar
Urzúa, Marcela
Rocha Martín, Javier
Fernández Lafuente, Roberto
author_role author
author2 Hocine, El Siar
Urzúa, Marcela
Rocha Martín, Javier
Fernández Lafuente, Roberto
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 577.1
577.15
547.94
544.478
Enzyme specificity
Conformational changes induced by immobilization
Enzyme tuning by immobilization
Bioquímica (Biología)
2403 Bioquímica
2302.09 Enzimología
2302.27 Proteínas
2210.01 Catálisis
topic 577.1
577.15
547.94
544.478
Enzyme specificity
Conformational changes induced by immobilization
Enzyme tuning by immobilization
Bioquímica (Biología)
2403 Bioquímica
2302.09 Enzimología
2302.27 Proteínas
2210.01 Catálisis
description Ficin extract has been immobilized using different supports: glyoxyl and Aspartic/1,6 hexamethylenediamine (Asp/HA) agarose beads. The latter was later submitted to glutaraldehyde modification to get covalent immobilization. The activities of these 3 kinds of biocatalysts were compared utilizing 4 different substrates, casein, hemoglobin and bovine serum albumin and benzoyl-arginine-p-nitroanilide at pH 7 and 5. Using glyoxyl-agarose, the effect of enzyme-support reaction time on the activity versus the four substrates at both pH values was studied. Reaction time has been shown to distort the enzyme due to an increase in the number of covalent support-enzyme bonds. Surprisingly, for all the substrates and conditions the prolongation of the enzyme-support reaction did not imply a decrease in enzyme activity. Using the Asp/HA supports (with different amount of HA) differences in the effect on enzyme activity versus the different substrates are much more significant, while with some substrates the immobilization produced a decrease in enzyme activity, with in other cases the activity increased. These different effects are even increased after glutaraldehyde treatment. That way, the conformational changes induced by the biocatalyst immobilization or the chemical modification fully altered the enzyme protein specificity. This may also have some implications when following enzyme inactivation.
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/20.500.14352/112066
url https://hdl.handle.net/20.500.14352/112066
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Ciencia e Innovación http://dx.doi.org/10.13039/501100004837 Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023 PID2022–136535OB-I00
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
_version_ 1869418262582788096
score 15,81155

Similares en LA Referencia