Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme

Detalhes bibliográficos
Autor(a) principal: Tavano, Olga Luisa
Data de Publicação: 2013
Outros Autores: Fernandez-Lafuente, Roberto, Goulart, Antonio José [UNESP], Monti, Rubens [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.procbio.2013.05.009
http://hdl.handle.net/11449/75776
Resumo: A simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U mg-1 protein and 7.03 U mg-1 protein for free and immobilized preparations, respectively). © 2013 Elsevier Ltd.
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spelling Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzymeAgaroseBeta-amylaseGlutaraldehydeImmobilizationSweet potatoBiocatalyst particleDiffusion limitationsGlutaraldehyde-agaroseGlutaraldehydesSimplified procedureAmylasesProteinsRadioactive waste vitrificationEnzyme immobilizationIpomoea batatasSolanum tuberosumA simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U mg-1 protein and 7.03 U mg-1 protein for free and immobilized preparations, respectively). © 2013 Elsevier Ltd.Department of Nutrition ICS Universidade Federal Do Triângulo Mineiro, Rua Getulio Guaritá, 159, Uberaba, MG, CEP 38025 360Department of Biocatalysis ICP-CSIC Campus UAM-CSIC, Cantoblanco, Madrid ZC 28049Department of Food Nutrition Faculty of Pharmaceutical Sciences UNESP-São Paulo State University, Rodovia Araraquara-Jaú, Km1, 14840-000, Araraquara, SPDepartment of Food Nutrition Faculty of Pharmaceutical Sciences UNESP-São Paulo State University, Rodovia Araraquara-Jaú, Km1, 14840-000, Araraquara, SPUniversidade Federal do Triângulo Mineiro (UFTM)ICP-CSICUniversidade Estadual Paulista (Unesp)Tavano, Olga LuisaFernandez-Lafuente, RobertoGoulart, Antonio José [UNESP]Monti, Rubens [UNESP]2014-05-27T11:29:49Z2014-05-27T11:29:49Z2013-07-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1054-1058application/pdfhttp://dx.doi.org/10.1016/j.procbio.2013.05.009Process Biochemistry, v. 48, n. 7, p. 1054-1058, 2013.1359-5113http://hdl.handle.net/11449/7577610.1016/j.procbio.2013.05.009WOS:0003229286000092-s2.0-848800506422-s2.0-84880050642.pdf5962867835836749Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengProcess Biochemistry2.6160,761info:eu-repo/semantics/openAccess2024-06-21T12:46:49Zoai:repositorio.unesp.br:11449/75776Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:00:03.954904Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
title Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
spellingShingle Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
Tavano, Olga Luisa
Agarose
Beta-amylase
Glutaraldehyde
Immobilization
Sweet potato
Biocatalyst particle
Diffusion limitations
Glutaraldehyde-agarose
Glutaraldehydes
Simplified procedure
Amylases
Proteins
Radioactive waste vitrification
Enzyme immobilization
Ipomoea batatas
Solanum tuberosum
title_short Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
title_full Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
title_fullStr Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
title_full_unstemmed Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
title_sort Optimization of the immobilization of sweet potato amylase using glutaraldehyde-agarose support. Characterization of the immobilized enzyme
author Tavano, Olga Luisa
author_facet Tavano, Olga Luisa
Fernandez-Lafuente, Roberto
Goulart, Antonio José [UNESP]
Monti, Rubens [UNESP]
author_role author
author2 Fernandez-Lafuente, Roberto
Goulart, Antonio José [UNESP]
Monti, Rubens [UNESP]
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade Federal do Triângulo Mineiro (UFTM)
ICP-CSIC
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Tavano, Olga Luisa
Fernandez-Lafuente, Roberto
Goulart, Antonio José [UNESP]
Monti, Rubens [UNESP]
dc.subject.por.fl_str_mv Agarose
Beta-amylase
Glutaraldehyde
Immobilization
Sweet potato
Biocatalyst particle
Diffusion limitations
Glutaraldehyde-agarose
Glutaraldehydes
Simplified procedure
Amylases
Proteins
Radioactive waste vitrification
Enzyme immobilization
Ipomoea batatas
Solanum tuberosum
topic Agarose
Beta-amylase
Glutaraldehyde
Immobilization
Sweet potato
Biocatalyst particle
Diffusion limitations
Glutaraldehyde-agarose
Glutaraldehydes
Simplified procedure
Amylases
Proteins
Radioactive waste vitrification
Enzyme immobilization
Ipomoea batatas
Solanum tuberosum
description A simplified procedure for the preparation of immobilized beta-amylase using non-purified extract from fresh sweet potato tubers is established in this paper, using differently activated agarose supports. Beta-amylase glutaraldehyde derivative was the preparation with best features, presenting improved temperature and pH stability and activity. The possibility of reusing the amylase was also shown, when this immobilized enzyme was fully active for five cycles of use. However, immobilization decreased enzyme activity to around 15%. This seems to be mainly due to diffusion limitations of the starch inside the pores of the biocatalyst particles. A fifteen-fold increase in the Km was noticed, while the decrease of Vmax was only 30% (10.1 U mg-1 protein and 7.03 U mg-1 protein for free and immobilized preparations, respectively). © 2013 Elsevier Ltd.
publishDate 2013
dc.date.none.fl_str_mv 2013-07-01
2014-05-27T11:29:49Z
2014-05-27T11:29:49Z
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/10.1016/j.procbio.2013.05.009
Process Biochemistry, v. 48, n. 7, p. 1054-1058, 2013.
1359-5113
http://hdl.handle.net/11449/75776
10.1016/j.procbio.2013.05.009
WOS:000322928600009
2-s2.0-84880050642
2-s2.0-84880050642.pdf
5962867835836749
url http://dx.doi.org/10.1016/j.procbio.2013.05.009
http://hdl.handle.net/11449/75776
identifier_str_mv Process Biochemistry, v. 48, n. 7, p. 1054-1058, 2013.
1359-5113
10.1016/j.procbio.2013.05.009
WOS:000322928600009
2-s2.0-84880050642
2-s2.0-84880050642.pdf
5962867835836749
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Process Biochemistry
2.616
0,761
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 1054-1058
application/pdf
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
_version_ 1808128734583062528

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