Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking

Ficin fully immobilized on Asp-agarose beads at pH 7 but not on an aminated support. This made enzyme adsorption plus glutaraldehyde modification non-viable for this enzyme. Modifying glyoxyl-agarose beads with mixtures of Asp and 1,6-hexamethylenediamine (HA) at different ratios, mixed anion/cation...

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Autores: Gonzalez Vasquez, Alex D., Hocine, El Siar, Alcántara, Andrés R., Urzúa, Marcela, Rocha Martín, Javier, Fernandez 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:20.500.14352/112059
Acceso en línea:https://hdl.handle.net/20.500.14352/112059
Access Level:acceso abierto
Palabra clave:577.15
66.098
Ficin immobilization-stabilization
Glutaraldehyde
Heterofunctional supports
Biología molecular (Biología)
Bioquímica (Biología)
2415 Biología Molecular
2403 Bioquímica
2302.09 Enzimología
3302.90 Ingeniería Bioquímica
id ES_8bcdee8d1244d8034c5510bdc1d463b7
oai_identifier_str oai:docta.ucm.es:20.500.14352/112059
network_acronym_str ES
network_name_str España
repository_id_str
spelling Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinkingGonzalez Vasquez, Alex D.Hocine, El SiarAlcántara, Andrés R.Urzúa, MarcelaRocha Martín, JavierFernandez Lafuente, Roberto577.1566.098Ficin immobilization-stabilizationGlutaraldehydeHeterofunctional supportsBiología molecular (Biología)Bioquímica (Biología)2415 Biología Molecular2403 Bioquímica2302.09 Enzimología3302.90 Ingeniería BioquímicaFicin fully immobilized on Asp-agarose beads at pH 7 but not on an aminated support. This made enzyme adsorption plus glutaraldehyde modification non-viable for this enzyme. Modifying glyoxyl-agarose beads with mixtures of Asp and 1,6-hexamethylenediamine (HA) at different ratios, mixed anion/cation exchanger supports were built. Only if HA greatly exceed Asp in the support, immobilization did not work. While only using the Asp-agarose support immobilized enzyme molecules were only ionically adsorbed after glutaraldehyde treatment (visualized in SDS-PAGE analysis), the mixed supports gave covalent immobilization. The glutaraldehyde modification of these biocatalysts permitted to establish covalent bonds with the support, and this was more effective when using higher amounts of HA in the support. When around 60 % of the groups in the support were HA, the treatment with glutaraldehyde fully suppressed enzyme release from the support after boiling in SDS. The glutaraldehyde treated biocatalysts were more stable than just the adsorbed enzymes or the enzyme adsorbed only on Asp supports and then treated with glutaraldehyde (the optimal biocatalyst retained 90 % of the initial activity while the just adsorbed ficin retained 50 % of the initial activity). This strategy can be utilized to immobilize other proteins with high isoelectric points following this immobilization strategy.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/20.500.14352/112059reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)InglésengMICIN Not available 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:20.500.14352/1120592026-06-02T12:44:21Z
dc.title.none.fl_str_mv Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
title Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
spellingShingle Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
Gonzalez Vasquez, Alex D.
577.15
66.098
Ficin immobilization-stabilization
Glutaraldehyde
Heterofunctional supports
Biología molecular (Biología)
Bioquímica (Biología)
2415 Biología Molecular
2403 Bioquímica
2302.09 Enzimología
3302.90 Ingeniería Bioquímica
title_short Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
title_full Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
title_fullStr Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
title_full_unstemmed Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
title_sort Designing mixed cationic/anionic supports to covalently immobilize/stabilize enzymes with high isoelectric point by enzyme adsorption and support-enzyme glutaraldehyde crosslinking
dc.creator.none.fl_str_mv Gonzalez Vasquez, Alex D.
Hocine, El Siar
Alcántara, Andrés R.
Urzúa, Marcela
Rocha Martín, Javier
Fernandez Lafuente, Roberto
author Gonzalez Vasquez, Alex D.
author_facet Gonzalez Vasquez, Alex D.
Hocine, El Siar
Alcántara, Andrés R.
Urzúa, Marcela
Rocha Martín, Javier
Fernandez Lafuente, Roberto
author_role author
author2 Hocine, El Siar
Alcántara, Andrés R.
Urzúa, Marcela
Rocha Martín, Javier
Fernandez 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 577.15
66.098
Ficin immobilization-stabilization
Glutaraldehyde
Heterofunctional supports
Biología molecular (Biología)
Bioquímica (Biología)
2415 Biología Molecular
2403 Bioquímica
2302.09 Enzimología
3302.90 Ingeniería Bioquímica
topic 577.15
66.098
Ficin immobilization-stabilization
Glutaraldehyde
Heterofunctional supports
Biología molecular (Biología)
Bioquímica (Biología)
2415 Biología Molecular
2403 Bioquímica
2302.09 Enzimología
3302.90 Ingeniería Bioquímica
description Ficin fully immobilized on Asp-agarose beads at pH 7 but not on an aminated support. This made enzyme adsorption plus glutaraldehyde modification non-viable for this enzyme. Modifying glyoxyl-agarose beads with mixtures of Asp and 1,6-hexamethylenediamine (HA) at different ratios, mixed anion/cation exchanger supports were built. Only if HA greatly exceed Asp in the support, immobilization did not work. While only using the Asp-agarose support immobilized enzyme molecules were only ionically adsorbed after glutaraldehyde treatment (visualized in SDS-PAGE analysis), the mixed supports gave covalent immobilization. The glutaraldehyde modification of these biocatalysts permitted to establish covalent bonds with the support, and this was more effective when using higher amounts of HA in the support. When around 60 % of the groups in the support were HA, the treatment with glutaraldehyde fully suppressed enzyme release from the support after boiling in SDS. The glutaraldehyde treated biocatalysts were more stable than just the adsorbed enzymes or the enzyme adsorbed only on Asp supports and then treated with glutaraldehyde (the optimal biocatalyst retained 90 % of the initial activity while the just adsorbed ficin retained 50 % of the initial activity). This strategy can be utilized to immobilize other proteins with high isoelectric points following this immobilization strategy.
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/20.500.14352/112059
url https://hdl.handle.net/20.500.14352/112059
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv MICIN Not available 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
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score 15,811543

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