Immobilization and Characterization of L-Asparaginase over Carbon Xerogels
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.3390/biotech11020010 http://hdl.handle.net/11449/239961 |
Resumo: | L-asparaginase (ASNase) is an aminohydrolase currently used in the pharmaceutical and food industries. Enzyme immobilization is an exciting option for both applications, allowing for a more straightforward recovery and increased stability. High surface area and customizable porosity make carbon xerogels (CXs) promising materials for ASNase immobilization. This work describes the influence of contact time, pH, and ASNase concentration on the immobilization yield (IY) and relative recovered activity (RRA) using the Central Composite Design methodology. The most promising results were obtained using CX with an average pore size of 4 nm (CX-4), reaching IY and RRA of 100%. At the optimal conditions (contact time 49 min, pH 6.73, and [ASNase] 0.26 mg·mL−1), the ASNase-CXs biocomposite was characterized and evaluated in terms of kinetic properties and operational, thermal, and pH stabilities. The immobilized ASNase onto CX-4 retained 71% of its original activity after six continuous reaction cycles, showed good thermal stability at 37 °C (RRA of 91% after 90 min), and was able to adapt to both acidic and alkaline environments. Finally, the results indicated a 3.9-fold increase in the immobilized ASNase affinity for the substrate, confirming the potential of CXs as a support for ASNase and as a cost-effective tool for subsequent use in the therapeutic and food sectors. |
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Immobilization and Characterization of L-Asparaginase over Carbon Xerogelscarbon xerogelscentral composite designenzyme immobilizationL-asparaginasephysical adsorptionL-asparaginase (ASNase) is an aminohydrolase currently used in the pharmaceutical and food industries. Enzyme immobilization is an exciting option for both applications, allowing for a more straightforward recovery and increased stability. High surface area and customizable porosity make carbon xerogels (CXs) promising materials for ASNase immobilization. This work describes the influence of contact time, pH, and ASNase concentration on the immobilization yield (IY) and relative recovered activity (RRA) using the Central Composite Design methodology. The most promising results were obtained using CX with an average pore size of 4 nm (CX-4), reaching IY and RRA of 100%. At the optimal conditions (contact time 49 min, pH 6.73, and [ASNase] 0.26 mg·mL−1), the ASNase-CXs biocomposite was characterized and evaluated in terms of kinetic properties and operational, thermal, and pH stabilities. The immobilized ASNase onto CX-4 retained 71% of its original activity after six continuous reaction cycles, showed good thermal stability at 37 °C (RRA of 91% after 90 min), and was able to adapt to both acidic and alkaline environments. Finally, the results indicated a 3.9-fold increase in the immobilized ASNase affinity for the substrate, confirming the potential of CXs as a support for ASNase and as a cost-effective tool for subsequent use in the therapeutic and food sectors.Federación Española de Enfermedades RarasConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação para a Ciência e a TecnologiaLSRE-LCM—Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials Faculty of Engineering University of PortoALiCE—Associate Laboratory in Chemical Engineering Faculty of Engineering University of PortoLAQV-REQUIMTE Department of Chemistry NOVA School of Science and Technology Universidade NOVA de LisboaCICECO-Aveiro Institute of Materials Department of Chemistry University of AveiroDepartment of Engineering Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP-University Estadual PaulistaDepartment of Engineering Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP-University Estadual PaulistaCNPq: 312463/2021-9Fundação para a Ciência e a Tecnologia: CEECINST/00102/2018University of PortoUniversidade NOVA de LisboaUniversity of AveiroUniversidade Estadual Paulista (UNESP)Barros, Rita A. M.Cristóvão, Raquel O.Carabineiro, Sónia A. C.Neves, Márcia C.Freire, Mara G.Faria, Joaquim L.Santos-Ebinuma, Valéria C. [UNESP]Tavares, Ana P. M.Silva, Cláudia G.2023-03-01T19:55:12Z2023-03-01T19:55:12Z2022-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.3390/biotech11020010BioTech, v. 11, n. 2, 2022.2673-6284http://hdl.handle.net/11449/23996110.3390/biotech110200102-s2.0-85129151854Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBioTechinfo:eu-repo/semantics/openAccess2023-03-01T19:55:12Zoai:repositorio.unesp.br:11449/239961Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-03-01T19:55:12Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
title |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
spellingShingle |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels Barros, Rita A. M. carbon xerogels central composite design enzyme immobilization L-asparaginase physical adsorption |
title_short |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
title_full |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
title_fullStr |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
title_full_unstemmed |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
title_sort |
Immobilization and Characterization of L-Asparaginase over Carbon Xerogels |
author |
Barros, Rita A. M. |
author_facet |
Barros, Rita A. M. Cristóvão, Raquel O. Carabineiro, Sónia A. C. Neves, Márcia C. Freire, Mara G. Faria, Joaquim L. Santos-Ebinuma, Valéria C. [UNESP] Tavares, Ana P. M. Silva, Cláudia G. |
author_role |
author |
author2 |
Cristóvão, Raquel O. Carabineiro, Sónia A. C. Neves, Márcia C. Freire, Mara G. Faria, Joaquim L. Santos-Ebinuma, Valéria C. [UNESP] Tavares, Ana P. M. Silva, Cláudia G. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
University of Porto Universidade NOVA de Lisboa University of Aveiro Universidade Estadual Paulista (UNESP) |
dc.contributor.author.fl_str_mv |
Barros, Rita A. M. Cristóvão, Raquel O. Carabineiro, Sónia A. C. Neves, Márcia C. Freire, Mara G. Faria, Joaquim L. Santos-Ebinuma, Valéria C. [UNESP] Tavares, Ana P. M. Silva, Cláudia G. |
dc.subject.por.fl_str_mv |
carbon xerogels central composite design enzyme immobilization L-asparaginase physical adsorption |
topic |
carbon xerogels central composite design enzyme immobilization L-asparaginase physical adsorption |
description |
L-asparaginase (ASNase) is an aminohydrolase currently used in the pharmaceutical and food industries. Enzyme immobilization is an exciting option for both applications, allowing for a more straightforward recovery and increased stability. High surface area and customizable porosity make carbon xerogels (CXs) promising materials for ASNase immobilization. This work describes the influence of contact time, pH, and ASNase concentration on the immobilization yield (IY) and relative recovered activity (RRA) using the Central Composite Design methodology. The most promising results were obtained using CX with an average pore size of 4 nm (CX-4), reaching IY and RRA of 100%. At the optimal conditions (contact time 49 min, pH 6.73, and [ASNase] 0.26 mg·mL−1), the ASNase-CXs biocomposite was characterized and evaluated in terms of kinetic properties and operational, thermal, and pH stabilities. The immobilized ASNase onto CX-4 retained 71% of its original activity after six continuous reaction cycles, showed good thermal stability at 37 °C (RRA of 91% after 90 min), and was able to adapt to both acidic and alkaline environments. Finally, the results indicated a 3.9-fold increase in the immobilized ASNase affinity for the substrate, confirming the potential of CXs as a support for ASNase and as a cost-effective tool for subsequent use in the therapeutic and food sectors. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-06-01 2023-03-01T19:55:12Z 2023-03-01T19:55:12Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.3390/biotech11020010 BioTech, v. 11, n. 2, 2022. 2673-6284 http://hdl.handle.net/11449/239961 10.3390/biotech11020010 2-s2.0-85129151854 |
url |
http://dx.doi.org/10.3390/biotech11020010 http://hdl.handle.net/11449/239961 |
identifier_str_mv |
BioTech, v. 11, n. 2, 2022. 2673-6284 10.3390/biotech11020010 2-s2.0-85129151854 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
BioTech |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1797789874228559872 |