A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da FURG (RI FURG) |
Texto Completo: | http://repositorio.furg.br/handle/1/4676 http://dx.doi.org/10.5935/0100-4042.20140128 |
Resumo: | This study aimed to evaluate B-galactosidase immobilization. For this purpose, the ionic strength of the buffer, reaction time, amount of the immobilization support, and pH were evaluated by a central composite design. Assay 8, which consisted of 1.5 mol L −1 phosphate buffer (pH 7.5) and a reaction time of 2 h, produced the maximum yield. Eupergit® C (400 mg) was subsequently use as an immobilization support. Immobilization kinetics wereinvestigated, and a significant increase in the yield was obtained after immobilization compared with that obtained from assay 8 (22.0 U mL −1 vs. 15.6 U mL −1). The enzyme efficiency of actuation was evaluated using o -nitrophenyl-B-d-galactopyranoside and lactose, with lactose providing better results. The reuse of B-galactosidase was evaluated, and more than 50% of the initial enzyme activity was maintained after five cycles of use. Enzyme characterization revealed that immobilization improved some aspects of the thermostability of B-galactosidase. |
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Braga, Anna Rafaela CavalcanteSilva, Marceli FernandesOliveira, José Vladimir deTreichel, HelenKalil, Susana Juliano2014-11-22T22:05:37Z2014-11-22T22:05:37Z2014BRAGA, Anna Rafaela Cavalcante et al. A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization. Química Nova, v. 37, n. 5, p. 796-803, 2014. Diponível em: <http://quimicanova.sbq.org.br/imagebank/pdf/v37n5a07.pdf>. Acesso em: 31 out. 2014.1678-7064http://repositorio.furg.br/handle/1/4676http://dx.doi.org/10.5935/0100-4042.20140128This study aimed to evaluate B-galactosidase immobilization. For this purpose, the ionic strength of the buffer, reaction time, amount of the immobilization support, and pH were evaluated by a central composite design. Assay 8, which consisted of 1.5 mol L −1 phosphate buffer (pH 7.5) and a reaction time of 2 h, produced the maximum yield. Eupergit® C (400 mg) was subsequently use as an immobilization support. Immobilization kinetics wereinvestigated, and a significant increase in the yield was obtained after immobilization compared with that obtained from assay 8 (22.0 U mL −1 vs. 15.6 U mL −1). The enzyme efficiency of actuation was evaluated using o -nitrophenyl-B-d-galactopyranoside and lactose, with lactose providing better results. The reuse of B-galactosidase was evaluated, and more than 50% of the initial enzyme activity was maintained after five cycles of use. Enzyme characterization revealed that immobilization improved some aspects of the thermostability of B-galactosidase.engBinding kineticsB-galactosidaseScanning electron microscopyA new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterizationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FURG (RI FURG)instname:Universidade Federal do Rio Grande (FURG)instacron:FURGORIGINALA NEW APPROACH TO EVALUATE IMMOBILIZATION OF β-GALACTOSIDASE ON EUPERGIT C.pdfA NEW APPROACH TO EVALUATE IMMOBILIZATION OF β-GALACTOSIDASE ON EUPERGIT C.pdfapplication/pdf720512https://repositorio.furg.br/bitstream/1/4676/1/A%20NEW%20APPROACH%20TO%20EVALUATE%20IMMOBILIZATION%20OF%20%ce%b2-GALACTOSIDASE%20ON%20EUPERGIT%20C.pdff27890ceed737b9a7e7e76bdcabb6693MD51open accessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.furg.br/bitstream/1/4676/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52open access1/46762014-11-22 20:05:37.815open accessoai:repositorio.furg.br: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Repositório InstitucionalPUBhttps://repositorio.furg.br/oai/request || http://200.19.254.174/oai/requestopendoar:2014-11-22T22:05:37Repositório Institucional da FURG (RI FURG) - Universidade Federal do Rio Grande (FURG)false |
dc.title.pt_BR.fl_str_mv |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
title |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
spellingShingle |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization Braga, Anna Rafaela Cavalcante Binding kinetics B-galactosidase Scanning electron microscopy |
title_short |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
title_full |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
title_fullStr |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
title_full_unstemmed |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
title_sort |
A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization |
author |
Braga, Anna Rafaela Cavalcante |
author_facet |
Braga, Anna Rafaela Cavalcante Silva, Marceli Fernandes Oliveira, José Vladimir de Treichel, Helen Kalil, Susana Juliano |
author_role |
author |
author2 |
Silva, Marceli Fernandes Oliveira, José Vladimir de Treichel, Helen Kalil, Susana Juliano |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Braga, Anna Rafaela Cavalcante Silva, Marceli Fernandes Oliveira, José Vladimir de Treichel, Helen Kalil, Susana Juliano |
dc.subject.por.fl_str_mv |
Binding kinetics B-galactosidase Scanning electron microscopy |
topic |
Binding kinetics B-galactosidase Scanning electron microscopy |
description |
This study aimed to evaluate B-galactosidase immobilization. For this purpose, the ionic strength of the buffer, reaction time, amount of the immobilization support, and pH were evaluated by a central composite design. Assay 8, which consisted of 1.5 mol L −1 phosphate buffer (pH 7.5) and a reaction time of 2 h, produced the maximum yield. Eupergit® C (400 mg) was subsequently use as an immobilization support. Immobilization kinetics wereinvestigated, and a significant increase in the yield was obtained after immobilization compared with that obtained from assay 8 (22.0 U mL −1 vs. 15.6 U mL −1). The enzyme efficiency of actuation was evaluated using o -nitrophenyl-B-d-galactopyranoside and lactose, with lactose providing better results. The reuse of B-galactosidase was evaluated, and more than 50% of the initial enzyme activity was maintained after five cycles of use. Enzyme characterization revealed that immobilization improved some aspects of the thermostability of B-galactosidase. |
publishDate |
2014 |
dc.date.accessioned.fl_str_mv |
2014-11-22T22:05:37Z |
dc.date.available.fl_str_mv |
2014-11-22T22:05:37Z |
dc.date.issued.fl_str_mv |
2014 |
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.citation.fl_str_mv |
BRAGA, Anna Rafaela Cavalcante et al. A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization. Química Nova, v. 37, n. 5, p. 796-803, 2014. Diponível em: <http://quimicanova.sbq.org.br/imagebank/pdf/v37n5a07.pdf>. Acesso em: 31 out. 2014. |
dc.identifier.uri.fl_str_mv |
http://repositorio.furg.br/handle/1/4676 |
dc.identifier.issn.none.fl_str_mv |
1678-7064 |
dc.identifier.doi.pt_BR.fl_str_mv |
http://dx.doi.org/10.5935/0100-4042.20140128 |
identifier_str_mv |
BRAGA, Anna Rafaela Cavalcante et al. A new approach to evaluate immobilization of B-galactosidase on eupergit c: structural, kinetic, and thermal characterization. Química Nova, v. 37, n. 5, p. 796-803, 2014. Diponível em: <http://quimicanova.sbq.org.br/imagebank/pdf/v37n5a07.pdf>. Acesso em: 31 out. 2014. 1678-7064 |
url |
http://repositorio.furg.br/handle/1/4676 http://dx.doi.org/10.5935/0100-4042.20140128 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da FURG (RI FURG) instname:Universidade Federal do Rio Grande (FURG) instacron:FURG |
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Universidade Federal do Rio Grande (FURG) |
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FURG |
institution |
FURG |
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Repositório Institucional da FURG (RI FURG) |
collection |
Repositório Institucional da FURG (RI FURG) |
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