Purification and characterization of an a-galactosidase from Aspergillus fumigatus

Aspergillus fumigatus secreted invertase (b-fructofuranosidase) and a-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). a-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtra...

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Detalhes bibliográficos
Autores: Rezende, Sebastião Tavares de, Guimarães, Valéria Monteze, Rodrigues, Marília de Castro, Felix, Carlos Roberto
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2005
País:Brasil
Recursos:Universidade Federal de Viçosa (UFV)
Repositório:LOCUS Repositório Institucional da UFV
Idioma:inglês
OAI Identifier:oai:locus.ufv.br:123456789/25582
Acesso em linha:http://dx.doi.org/10.1590/S1516-89132005000200005
http://www.locus.ufv.br/handle/123456789/25582
Access Level:Acceso aberto
Palavra-chave:Aspergillus fumigatus
a-galactosidase
Raffinose oligosaccharides
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spelling Purification and characterization of an a-galactosidase from Aspergillus fumigatusAspergillus fumigatusa-galactosidaseRaffinose oligosaccharidesAspergillus fumigatus secreted invertase (b-fructofuranosidase) and a-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). a-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtration and two ion exchange chromatographies and showed Mw of 54.7 kDa. The purified enzyme showed maximal activity against p-nitrophenyl-a-D-galactopyranoside (pNPGal) at pH 4.5-5.5 and 55 °C, and retained about 80% of the original activity after incubation for 90 minutes at 50ºC. The KM for pNPGal was 0.3 mM. Melibiose was hydrolyzed by the enzyme but raffinose was very poor substrate.O fungo termofílico Aspergillus fumigatus secreta as enzimas invertase (b-frutofuranosidase) e a-galactosidase (a-D-galactosídeo galactohi-drolase) que estão envolvidas na hidrólise completa dos oligossacarídeos de rafinose. A enzima a-galactosidase foi produzida em meio de cultura do fungo Aspergillus fumigatus crescido por 36 h a 42 °C em meio mineral mínimo contendo os açúcares galactose, ou melibiose, ou rafinose como fontes de carbono. A enzima foi purificada por filtração em gel, seguida por duas cromatografias de troca iônica. A massa molecular da a-galactosidase determinada por SDS-PAGE foi de 54,7 kDa. A atividade máxima da enzima purificada, utilizando o substrato r-nitrofenil-a-D-galactopiranosídeo (rNPGal) foi na faixa de pH entre 4,5 e 5,5 e a 55 °C. A enzima manteve aproximadamente 80% de sua atividade original mesmo após pré-incubação por 90 minutos a 50 °C. O valor de KM para o substrato rNPGal foi 0,3 mM. A enzima foi capaz de hidrolisar melibiose, mas sua atividade foi muito reduzida na presença do substrato rafinose.Brazilian Archives of Biology and Technology2019-05-29T11:30:34Z2019-05-29T11:30:34Z2005-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlepdfapplication/pdf1678-4324http://dx.doi.org/10.1590/S1516-89132005000200005http://www.locus.ufv.br/handle/123456789/25582engv. 48, n. 02, p. 195-202, mar. 2005info:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVRezende, Sebastião Tavares deGuimarães, Valéria MontezeRodrigues, Marília de CastroFelix, Carlos Roberto2024-07-12T08:26:29Zoai:locus.ufv.br:123456789/25582Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452024-07-12T08:26:29LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.none.fl_str_mv Purification and characterization of an a-galactosidase from Aspergillus fumigatus
title Purification and characterization of an a-galactosidase from Aspergillus fumigatus
spellingShingle Purification and characterization of an a-galactosidase from Aspergillus fumigatus
Rezende, Sebastião Tavares de
Aspergillus fumigatus
a-galactosidase
Raffinose oligosaccharides
title_short Purification and characterization of an a-galactosidase from Aspergillus fumigatus
title_full Purification and characterization of an a-galactosidase from Aspergillus fumigatus
title_fullStr Purification and characterization of an a-galactosidase from Aspergillus fumigatus
title_full_unstemmed Purification and characterization of an a-galactosidase from Aspergillus fumigatus
title_sort Purification and characterization of an a-galactosidase from Aspergillus fumigatus
dc.creator.none.fl_str_mv Rezende, Sebastião Tavares de
Guimarães, Valéria Monteze
Rodrigues, Marília de Castro
Felix, Carlos Roberto
author Rezende, Sebastião Tavares de
author_facet Rezende, Sebastião Tavares de
Guimarães, Valéria Monteze
Rodrigues, Marília de Castro
Felix, Carlos Roberto
author_role author
author2 Guimarães, Valéria Monteze
Rodrigues, Marília de Castro
Felix, Carlos Roberto
author2_role author
author
author
dc.subject.por.fl_str_mv Aspergillus fumigatus
a-galactosidase
Raffinose oligosaccharides
topic Aspergillus fumigatus
a-galactosidase
Raffinose oligosaccharides
description Aspergillus fumigatus secreted invertase (b-fructofuranosidase) and a-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). a-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtration and two ion exchange chromatographies and showed Mw of 54.7 kDa. The purified enzyme showed maximal activity against p-nitrophenyl-a-D-galactopyranoside (pNPGal) at pH 4.5-5.5 and 55 °C, and retained about 80% of the original activity after incubation for 90 minutes at 50ºC. The KM for pNPGal was 0.3 mM. Melibiose was hydrolyzed by the enzyme but raffinose was very poor substrate.
publishDate 2005
dc.date.none.fl_str_mv 2005-03
2019-05-29T11:30:34Z
2019-05-29T11:30:34Z
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 1678-4324
http://dx.doi.org/10.1590/S1516-89132005000200005
http://www.locus.ufv.br/handle/123456789/25582
identifier_str_mv 1678-4324
url http://dx.doi.org/10.1590/S1516-89132005000200005
http://www.locus.ufv.br/handle/123456789/25582
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv v. 48, n. 02, p. 195-202, mar. 2005
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv pdf
application/pdf
dc.publisher.none.fl_str_mv Brazilian Archives of Biology and Technology
publisher.none.fl_str_mv Brazilian Archives of Biology and Technology
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
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score 15.301629

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