Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles

This work evaluates different dendrimer-silica supports for the immobilization of enzymes by multipoint covalent binding. Thermolysin was immobilized on two dendrimers (PAMAM and carbosilane) with two different generations (zero (G0) and first (G1)). Results were compared with a control, a silica su...

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Autores: Hernández Corroto, Ester|||0000-0002-6001-3634, Sánchez Milla, María, Sánchez-Nieves Fernández, Javier|||0000-0003-0410-2285, Mata de la Mata, Francisco Javier de la|||0000-0003-0418-3935, Marina Alegre, María Luisa|||0000-0002-5583-1624, García López, María Concepción|||0000-0002-3383-6176
Tipo de recurso: artículo
Fecha de publicación:2020
País:España
Institución:Universidad de Alcalá (UAH)
Repositorio:e_Buah Biblioteca Digital Universidad de Alcalá
Idioma:inglés
OAI Identifier:oai:ebuah.uah.es:10017/59701
Acceso en línea:http://hdl.handle.net/10017/59701
https://dx.doi.org/10.1016/j.ijbiomac.2020.10.138
Access Level:acceso abierto
Palabra clave:Enzyme immobilization
Thermolysin
Carbosilane
PAMAM
Dendrimer
Silica
Química
Chemistry
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spelling Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cyclesHernández Corroto, Ester|||0000-0002-6001-3634Sánchez Milla, MaríaSánchez-Nieves Fernández, Javier|||0000-0003-0410-2285Mata de la Mata, Francisco Javier de la|||0000-0003-0418-3935Marina Alegre, María Luisa|||0000-0002-5583-1624García López, María Concepción|||0000-0002-3383-6176Enzyme immobilizationThermolysinCarbosilanePAMAMDendrimerSilicaQuímicaChemistryThis work evaluates different dendrimer-silica supports for the immobilization of enzymes by multipoint covalent binding. Thermolysin was immobilized on two dendrimers (PAMAM and carbosilane) with two different generations (zero (G0) and first (G1)). Results were compared with a control, a silica support functionalized with amonofunctional molecule. Dendrimers increased the number of available sites to bind the enzyme. Despite the enzyme was immobilized on all supports, G0 dendrimers immobilized a 30% more enzyme than G1. Thermolysin immobilized on G0 dendrimer supports showed the highest activity and could be employed in three consecutive hydrolysis cycles. Optimal immobilization time was 1 h while optimal protein loading was 25 mg enzyme/100 mg support. Enzyme activity was promoted when using 5 mg of immobilized enzyme at 750 rpm, 60 degrees C, and 2 h of hydrolysis. Under these conditions, the activity of thermolysin increased up to the 78% of the free enzyme activity. Kinetics of the hydrolysis reaction using the immobilized thermolysin was also studied and compared with the obtained using the free thermolysin. The addition of ZnCl2 and NaCl during the immobilization procedure increased thermolysin activity in the second (22% more) and in the third (14% more) hydrolysis clycles.Universidad de AlcaláMinisterio de Economía, Industria y CompetitividadComunidad de MadridCentro de Investigación Biomédica en Red en el área temática de Bioingeniería, Biomateriales y Nanomedicina20202020-12-15journal articlehttp://purl.org/coar/resource_type/c_6501NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10017/59701https://dx.doi.org/10.1016/j.ijbiomac.2020.10.138reponame:e_Buah Biblioteca Digital Universidad de Alcaláinstname:Universidad de Alcalá (UAH)InglésengMinisterio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 Not available CTQ2017-86224-PMinisterio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 Not available AGL2016-79010-RComunidad de Madrid http://dx.doi.org/10.13039/100012818 Not available S2013%2FABI-3028open accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:ebuah.uah.es:10017/597012026-06-18T11:13:07Z
dc.title.none.fl_str_mv Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
title Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
spellingShingle Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
Hernández Corroto, Ester|||0000-0002-6001-3634
Enzyme immobilization
Thermolysin
Carbosilane
PAMAM
Dendrimer
Silica
Química
Chemistry
title_short Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
title_full Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
title_fullStr Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
title_full_unstemmed Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
title_sort Immobilization of thermolysin enzyme on dendronized silica supports. Evaluation of its feasibility on multiple protein hydrolysis cycles
dc.creator.none.fl_str_mv Hernández Corroto, Ester|||0000-0002-6001-3634
Sánchez Milla, María
Sánchez-Nieves Fernández, Javier|||0000-0003-0410-2285
Mata de la Mata, Francisco Javier de la|||0000-0003-0418-3935
Marina Alegre, María Luisa|||0000-0002-5583-1624
García López, María Concepción|||0000-0002-3383-6176
author Hernández Corroto, Ester|||0000-0002-6001-3634
author_facet Hernández Corroto, Ester|||0000-0002-6001-3634
Sánchez Milla, María
Sánchez-Nieves Fernández, Javier|||0000-0003-0410-2285
Mata de la Mata, Francisco Javier de la|||0000-0003-0418-3935
Marina Alegre, María Luisa|||0000-0002-5583-1624
García López, María Concepción|||0000-0002-3383-6176
author_role author
author2 Sánchez Milla, María
Sánchez-Nieves Fernández, Javier|||0000-0003-0410-2285
Mata de la Mata, Francisco Javier de la|||0000-0003-0418-3935
Marina Alegre, María Luisa|||0000-0002-5583-1624
García López, María Concepción|||0000-0002-3383-6176
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Enzyme immobilization
Thermolysin
Carbosilane
PAMAM
Dendrimer
Silica
Química
Chemistry
topic Enzyme immobilization
Thermolysin
Carbosilane
PAMAM
Dendrimer
Silica
Química
Chemistry
description This work evaluates different dendrimer-silica supports for the immobilization of enzymes by multipoint covalent binding. Thermolysin was immobilized on two dendrimers (PAMAM and carbosilane) with two different generations (zero (G0) and first (G1)). Results were compared with a control, a silica support functionalized with amonofunctional molecule. Dendrimers increased the number of available sites to bind the enzyme. Despite the enzyme was immobilized on all supports, G0 dendrimers immobilized a 30% more enzyme than G1. Thermolysin immobilized on G0 dendrimer supports showed the highest activity and could be employed in three consecutive hydrolysis cycles. Optimal immobilization time was 1 h while optimal protein loading was 25 mg enzyme/100 mg support. Enzyme activity was promoted when using 5 mg of immobilized enzyme at 750 rpm, 60 degrees C, and 2 h of hydrolysis. Under these conditions, the activity of thermolysin increased up to the 78% of the free enzyme activity. Kinetics of the hydrolysis reaction using the immobilized thermolysin was also studied and compared with the obtained using the free thermolysin. The addition of ZnCl2 and NaCl during the immobilization procedure increased thermolysin activity in the second (22% more) and in the third (14% more) hydrolysis clycles.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-12-15
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10017/59701
https://dx.doi.org/10.1016/j.ijbiomac.2020.10.138
url http://hdl.handle.net/10017/59701
https://dx.doi.org/10.1016/j.ijbiomac.2020.10.138
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 Not available CTQ2017-86224-P
Ministerio de Economía y Competitividad http://dx.doi.org/10.13039/501100003329 Not available AGL2016-79010-R
Comunidad de Madrid http://dx.doi.org/10.13039/100012818 Not available S2013%2FABI-3028
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/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-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:e_Buah Biblioteca Digital Universidad de Alcalá
instname:Universidad de Alcalá (UAH)
instname_str Universidad de Alcalá (UAH)
reponame_str e_Buah Biblioteca Digital Universidad de Alcalá
collection e_Buah Biblioteca Digital Universidad de Alcalá
repository.name.fl_str_mv
repository.mail.fl_str_mv
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