Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques

Detalhes bibliográficos
Autor(a) principal: Silva, Carla J. S. M.
Data de Publicação: 2006
Outros Autores: Gübitz, Georg M., Paulo, Artur Cavaco
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/4401
Resumo: A full factorial design was used to study the influence of four different variables, namely polymer concentration, carbodiimide concentration, time of reaction and blocking agent concentration, on the coupling of a serine protease into a soluble–insoluble polymer (Eudragit S-100). All of the four factors studied have played a critical role in the protease coupling. Response surface methodology was used as an optimisation strategy to attain a conjugate with high activity yield and operational stability at 60 ◦C. Under optimised conditions (Eudragit, 2.5% w/v, carbodiimide, 0.2% w/v, coupling time, 1 h and blocking agent concentration, 0.05%), the conjugate activity yield was about 45% and its operational stability at 60 ◦C was increased by 1.7 times. After reusing the conjugate for five cycles, the remaining activity was still 72% of the initial value when compared with the native enzyme. Several tests confirmed that the enzyme was covalently crosslinked to Eudragit, which represents an improvement in the carbodiimide coupling of proteases into soluble–insoluble polymers.
id RCAP_0418953a0683787c5c9c385bf20a9e0f
oai_identifier_str oai:repositorium.sdum.uminho.pt:1822/4401
network_acronym_str RCAP
network_name_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository_id_str 7160
spelling Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniquesSerine proteasesEudragitCouplingFactorial designsScience & TechnologyA full factorial design was used to study the influence of four different variables, namely polymer concentration, carbodiimide concentration, time of reaction and blocking agent concentration, on the coupling of a serine protease into a soluble–insoluble polymer (Eudragit S-100). All of the four factors studied have played a critical role in the protease coupling. Response surface methodology was used as an optimisation strategy to attain a conjugate with high activity yield and operational stability at 60 ◦C. Under optimised conditions (Eudragit, 2.5% w/v, carbodiimide, 0.2% w/v, coupling time, 1 h and blocking agent concentration, 0.05%), the conjugate activity yield was about 45% and its operational stability at 60 ◦C was increased by 1.7 times. After reusing the conjugate for five cycles, the remaining activity was still 72% of the initial value when compared with the native enzyme. Several tests confirmed that the enzyme was covalently crosslinked to Eudragit, which represents an improvement in the carbodiimide coupling of proteases into soluble–insoluble polymers.Society of Chemical IndustryUniversidade do MinhoSilva, Carla J. S. M.Gübitz, Georg M.Paulo, Artur Cavaco20062006-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/4401engJournal of Chemical Technology and Biotechnology." ISSN 0268–2575. 81:8 (2006) 8-16.0268–257510.1002/jctb.1350info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:08:47Zoai:repositorium.sdum.uminho.pt:1822/4401Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:00:03.697278Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
title Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
spellingShingle Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
Silva, Carla J. S. M.
Serine proteases
Eudragit
Coupling
Factorial designs
Science & Technology
title_short Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
title_full Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
title_fullStr Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
title_full_unstemmed Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
title_sort Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
author Silva, Carla J. S. M.
author_facet Silva, Carla J. S. M.
Gübitz, Georg M.
Paulo, Artur Cavaco
author_role author
author2 Gübitz, Georg M.
Paulo, Artur Cavaco
author2_role author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Silva, Carla J. S. M.
Gübitz, Georg M.
Paulo, Artur Cavaco
dc.subject.por.fl_str_mv Serine proteases
Eudragit
Coupling
Factorial designs
Science & Technology
topic Serine proteases
Eudragit
Coupling
Factorial designs
Science & Technology
description A full factorial design was used to study the influence of four different variables, namely polymer concentration, carbodiimide concentration, time of reaction and blocking agent concentration, on the coupling of a serine protease into a soluble–insoluble polymer (Eudragit S-100). All of the four factors studied have played a critical role in the protease coupling. Response surface methodology was used as an optimisation strategy to attain a conjugate with high activity yield and operational stability at 60 ◦C. Under optimised conditions (Eudragit, 2.5% w/v, carbodiimide, 0.2% w/v, coupling time, 1 h and blocking agent concentration, 0.05%), the conjugate activity yield was about 45% and its operational stability at 60 ◦C was increased by 1.7 times. After reusing the conjugate for five cycles, the remaining activity was still 72% of the initial value when compared with the native enzyme. Several tests confirmed that the enzyme was covalently crosslinked to Eudragit, which represents an improvement in the carbodiimide coupling of proteases into soluble–insoluble polymers.
publishDate 2006
dc.date.none.fl_str_mv 2006
2006-01-01T00:00:00Z
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://hdl.handle.net/1822/4401
url http://hdl.handle.net/1822/4401
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Chemical Technology and Biotechnology." ISSN 0268–2575. 81:8 (2006) 8-16.
0268–2575
10.1002/jctb.1350
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Society of Chemical Industry
publisher.none.fl_str_mv Society of Chemical Industry
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
_version_ 1799132394403397632