Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications
Autor(a) principal: | |
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Data de Publicação: | 2008 |
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/1822/17298 |
Resumo: | Subtilisin E is an alkaline serine protease secreted by the Gram positive bacterium Bacillus subtilis and widely used in industry as a biocatalyst for various processes. The most common application of subtilisins is in laundry detergents. However, due to environmental concerns, the application of subtilisins to treat wool, is under study. There are some reports regarding the attempts to substitute the conventional chlorine treatment by an enzymatic process capable of providing the same characteristics to the fabric, like anti-shrinking and better uptake and fixation of the dyestuff. However, the degree of uncontrolled hydrolysis due to diffusion of the enzyme inside the wool fiber causes unacceptable losses of strength. To overcome this fact, and taking advantage of the x-ray crystallographic structure, the authors have modified subtilisin E genetically, increasing its molecular weight, to restrict the hydrolysis to the surface of the wool fibers. Therefore, three genetically modified enzymes with a molecular weight 2-fold to 4-fold higher than the native subtilisin E were produced and assessed for activity. The prokaryotic expression systems, pET25b (+), pET11b and pBAD C, were explored for the production of recombinant enzymes. The results demonstrated that regardless the expression system or strain used, chimeric subtilisins were not expressed with the correct folding. No active and soluble recombinant protein was recovered under the testing conditions. Despite this drawback, a novel approach was described to increase the molecular weight of subtilisin. The reported results are noteworthy and can indicate good guidelines for future work aiming at the solubilization of recombinant chimeric subtilisins. |
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Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applicationsIn vitro refoldingProtein engineeringSubtilisin EWool hydrolysisScience & TechnologySubtilisin E is an alkaline serine protease secreted by the Gram positive bacterium Bacillus subtilis and widely used in industry as a biocatalyst for various processes. The most common application of subtilisins is in laundry detergents. However, due to environmental concerns, the application of subtilisins to treat wool, is under study. There are some reports regarding the attempts to substitute the conventional chlorine treatment by an enzymatic process capable of providing the same characteristics to the fabric, like anti-shrinking and better uptake and fixation of the dyestuff. However, the degree of uncontrolled hydrolysis due to diffusion of the enzyme inside the wool fiber causes unacceptable losses of strength. To overcome this fact, and taking advantage of the x-ray crystallographic structure, the authors have modified subtilisin E genetically, increasing its molecular weight, to restrict the hydrolysis to the surface of the wool fibers. Therefore, three genetically modified enzymes with a molecular weight 2-fold to 4-fold higher than the native subtilisin E were produced and assessed for activity. The prokaryotic expression systems, pET25b (+), pET11b and pBAD C, were explored for the production of recombinant enzymes. The results demonstrated that regardless the expression system or strain used, chimeric subtilisins were not expressed with the correct folding. No active and soluble recombinant protein was recovered under the testing conditions. Despite this drawback, a novel approach was described to increase the molecular weight of subtilisin. The reported results are noteworthy and can indicate good guidelines for future work aiming at the solubilization of recombinant chimeric subtilisins.WileyUniversidade do MinhoAraújo, RitaPaulo, Artur CavacoCasal, Margarida20082008-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/17298eng1618-286310.1002/elsc.200700056http://onlinelibrary.wiley.com/doi/10.1002/elsc.200700056/abstractinfo: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:RCAAP2023-07-21T12:26:20Zoai:repositorium.sdum.uminho.pt:1822/17298Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:20:45.720021Repositório 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 |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
title |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
spellingShingle |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications Araújo, Rita In vitro refolding Protein engineering Subtilisin E Wool hydrolysis Science & Technology |
title_short |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
title_full |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
title_fullStr |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
title_full_unstemmed |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
title_sort |
Strategies towards the functionalization of subtilisin E from bacillus subtilis for wool finishing applications |
author |
Araújo, Rita |
author_facet |
Araújo, Rita Paulo, Artur Cavaco Casal, Margarida |
author_role |
author |
author2 |
Paulo, Artur Cavaco Casal, Margarida |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Araújo, Rita Paulo, Artur Cavaco Casal, Margarida |
dc.subject.por.fl_str_mv |
In vitro refolding Protein engineering Subtilisin E Wool hydrolysis Science & Technology |
topic |
In vitro refolding Protein engineering Subtilisin E Wool hydrolysis Science & Technology |
description |
Subtilisin E is an alkaline serine protease secreted by the Gram positive bacterium Bacillus subtilis and widely used in industry as a biocatalyst for various processes. The most common application of subtilisins is in laundry detergents. However, due to environmental concerns, the application of subtilisins to treat wool, is under study. There are some reports regarding the attempts to substitute the conventional chlorine treatment by an enzymatic process capable of providing the same characteristics to the fabric, like anti-shrinking and better uptake and fixation of the dyestuff. However, the degree of uncontrolled hydrolysis due to diffusion of the enzyme inside the wool fiber causes unacceptable losses of strength. To overcome this fact, and taking advantage of the x-ray crystallographic structure, the authors have modified subtilisin E genetically, increasing its molecular weight, to restrict the hydrolysis to the surface of the wool fibers. Therefore, three genetically modified enzymes with a molecular weight 2-fold to 4-fold higher than the native subtilisin E were produced and assessed for activity. The prokaryotic expression systems, pET25b (+), pET11b and pBAD C, were explored for the production of recombinant enzymes. The results demonstrated that regardless the expression system or strain used, chimeric subtilisins were not expressed with the correct folding. No active and soluble recombinant protein was recovered under the testing conditions. Despite this drawback, a novel approach was described to increase the molecular weight of subtilisin. The reported results are noteworthy and can indicate good guidelines for future work aiming at the solubilization of recombinant chimeric subtilisins. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008 2008-01-01T00:00:00Z |
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/1822/17298 |
url |
http://hdl.handle.net/1822/17298 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1618-2863 10.1002/elsc.200700056 http://onlinelibrary.wiley.com/doi/10.1002/elsc.200700056/abstract |
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 |
Wiley |
publisher.none.fl_str_mv |
Wiley |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799132671770624000 |