Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://dx.doi.org/10.1021/jf8030919 http://www.locus.ufv.br/handle/123456789/19852 |
Resumo: | Debaryomyces hansenii cells cultivated on galactose produced extracellular and intracellular α-galactosidases, which showed 54.5 and 54.8 kDa molecular mass (MALDI-TOF), 60 and 61 kDa (SDS−PAGE) and 5.15 and 4.15 pI values, respectively. The extracellular and intracellular deglycosylated forms presented 36 and 40 kDa molecular mass, with 40 and 34% carbohydrate content, respectively. The N-terminal sequences of the α-galactosidases were identical. Intracellular α-galactosidase showed smaller thermostability when compared to the extracellular enzyme. D. hansenii UFV-1 extracellular α-galactosidase presented higher kcat than the intracellular enzyme (7.16 vs 3.29 s^−1, respectively) for the p-nitrophenyl-α-d-galactopyranoside substrate. The Km for hydrolysis of pNPαGal, melibiose, stachyose, and raffinose were 0.32, 2.12, 10.8, and 32.8 mM, respectively. The intracellular enzyme was acompetitively inhibited by galactose (Ki = 0.70 mM), and it was inactivated by Cu(II) and Ag(I). Enzyme incubation with soy milk for 6 h at 55 °C reduced stachyose and raffinose amounts by 100 and 73%, respectively. |
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LOCUS Repositório Institucional da UFV |
repository_id_str |
2145 |
spelling |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzymeCharacterizationDebaryomyces hansenii UFV-1DeglycosylationGalacto-oligosaccharidesα-GalactosidasesDebaryomyces hansenii cells cultivated on galactose produced extracellular and intracellular α-galactosidases, which showed 54.5 and 54.8 kDa molecular mass (MALDI-TOF), 60 and 61 kDa (SDS−PAGE) and 5.15 and 4.15 pI values, respectively. The extracellular and intracellular deglycosylated forms presented 36 and 40 kDa molecular mass, with 40 and 34% carbohydrate content, respectively. The N-terminal sequences of the α-galactosidases were identical. Intracellular α-galactosidase showed smaller thermostability when compared to the extracellular enzyme. D. hansenii UFV-1 extracellular α-galactosidase presented higher kcat than the intracellular enzyme (7.16 vs 3.29 s^−1, respectively) for the p-nitrophenyl-α-d-galactopyranoside substrate. The Km for hydrolysis of pNPαGal, melibiose, stachyose, and raffinose were 0.32, 2.12, 10.8, and 32.8 mM, respectively. The intracellular enzyme was acompetitively inhibited by galactose (Ki = 0.70 mM), and it was inactivated by Cu(II) and Ag(I). Enzyme incubation with soy milk for 6 h at 55 °C reduced stachyose and raffinose amounts by 100 and 73%, respectively.Journal of Agricultural and Food Chemistry2018-05-29T10:24:11Z2018-05-29T10:24:11Z2009-02-18info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlepdfapplication/pdfhttp://dx.doi.org/10.1021/jf803091915205118http://www.locus.ufv.br/handle/123456789/19852engv. 57, n. 06, p. 2515–2522, Fevereiro 2009American Chemical Societyinfo:eu-repo/semantics/openAccessRezende, Sebastião T. deViana, Pollyanna A.Passos, Flávia Maria LopesOliveira, Jamil S.Teixeira, Kádima N.Santos, Alexandre M. C.Bemquerer, Marcelo P.Rosa, José C.Santoro, Marcelo M.Guimarães, Valéria M.reponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFV2024-07-12T06:50:21Zoai:locus.ufv.br:123456789/19852Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452024-07-12T06:50:21LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.none.fl_str_mv |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
title |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
spellingShingle |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme Rezende, Sebastião T. de Characterization Debaryomyces hansenii UFV-1 Deglycosylation Galacto-oligosaccharides α-Galactosidases |
title_short |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
title_full |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
title_fullStr |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
title_full_unstemmed |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
title_sort |
Debaryomyces hansenii UFV-1 intracellular α-Galactosidase characterization and comparative studies with the extracellular enzyme |
author |
Rezende, Sebastião T. de |
author_facet |
Rezende, Sebastião T. de Viana, Pollyanna A. Passos, Flávia Maria Lopes Oliveira, Jamil S. Teixeira, Kádima N. Santos, Alexandre M. C. Bemquerer, Marcelo P. Rosa, José C. Santoro, Marcelo M. Guimarães, Valéria M. |
author_role |
author |
author2 |
Viana, Pollyanna A. Passos, Flávia Maria Lopes Oliveira, Jamil S. Teixeira, Kádima N. Santos, Alexandre M. C. Bemquerer, Marcelo P. Rosa, José C. Santoro, Marcelo M. Guimarães, Valéria M. |
author2_role |
author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Rezende, Sebastião T. de Viana, Pollyanna A. Passos, Flávia Maria Lopes Oliveira, Jamil S. Teixeira, Kádima N. Santos, Alexandre M. C. Bemquerer, Marcelo P. Rosa, José C. Santoro, Marcelo M. Guimarães, Valéria M. |
dc.subject.por.fl_str_mv |
Characterization Debaryomyces hansenii UFV-1 Deglycosylation Galacto-oligosaccharides α-Galactosidases |
topic |
Characterization Debaryomyces hansenii UFV-1 Deglycosylation Galacto-oligosaccharides α-Galactosidases |
description |
Debaryomyces hansenii cells cultivated on galactose produced extracellular and intracellular α-galactosidases, which showed 54.5 and 54.8 kDa molecular mass (MALDI-TOF), 60 and 61 kDa (SDS−PAGE) and 5.15 and 4.15 pI values, respectively. The extracellular and intracellular deglycosylated forms presented 36 and 40 kDa molecular mass, with 40 and 34% carbohydrate content, respectively. The N-terminal sequences of the α-galactosidases were identical. Intracellular α-galactosidase showed smaller thermostability when compared to the extracellular enzyme. D. hansenii UFV-1 extracellular α-galactosidase presented higher kcat than the intracellular enzyme (7.16 vs 3.29 s^−1, respectively) for the p-nitrophenyl-α-d-galactopyranoside substrate. The Km for hydrolysis of pNPαGal, melibiose, stachyose, and raffinose were 0.32, 2.12, 10.8, and 32.8 mM, respectively. The intracellular enzyme was acompetitively inhibited by galactose (Ki = 0.70 mM), and it was inactivated by Cu(II) and Ag(I). Enzyme incubation with soy milk for 6 h at 55 °C reduced stachyose and raffinose amounts by 100 and 73%, respectively. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-02-18 2018-05-29T10:24:11Z 2018-05-29T10:24:11Z |
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/10.1021/jf8030919 15205118 http://www.locus.ufv.br/handle/123456789/19852 |
url |
http://dx.doi.org/10.1021/jf8030919 http://www.locus.ufv.br/handle/123456789/19852 |
identifier_str_mv |
15205118 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
v. 57, n. 06, p. 2515–2522, Fevereiro 2009 |
dc.rights.driver.fl_str_mv |
American Chemical Society info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
American Chemical Society |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
pdf application/pdf |
dc.publisher.none.fl_str_mv |
Journal of Agricultural and Food Chemistry |
publisher.none.fl_str_mv |
Journal of Agricultural and Food Chemistry |
dc.source.none.fl_str_mv |
reponame:LOCUS Repositório Institucional da UFV instname:Universidade Federal de Viçosa (UFV) instacron:UFV |
instname_str |
Universidade Federal de Viçosa (UFV) |
instacron_str |
UFV |
institution |
UFV |
reponame_str |
LOCUS Repositório Institucional da UFV |
collection |
LOCUS Repositório Institucional da UFV |
repository.name.fl_str_mv |
LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV) |
repository.mail.fl_str_mv |
fabiojreis@ufv.br |
_version_ |
1817559874415886336 |