Optimisation of a serine protease coupling to Eudragit S-100 by experimental design techniques
Autor(a) principal: | |
---|---|
Data de Publicação: | 2006 |
Outros Autores: | , |
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 |