Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii

Detalhes bibliográficos
Autor(a) principal: Schnitzhofer, W
Data de Publicação: 2007
Outros Autores: Weber, H.-J., Vršanská, M., Biely, P., Paulo, Artur Cavaco, Guebitz, G. M.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/13560
Resumo: Sclerotium rolfsii (strain CBS 350.80) was found to produce extraordinary high amounts of polygalacturonases (PGs). Two of these extracellular enzymes were purified by a recently introduced preparative electrophoretic device (isoelectric focusing mode of free flow electrophoresis). PG 1 (39.5 kDa, pI 6.5) and PG 2 (38 kDa, pI 5.4) exhibited quite similar properties, they were found to be both endo-acting enzymes. Both PGs cleaved penta- and trigalacturonic acid while tetragalacturonic acid was only cleaved when trigalacturonic acid was present. The latter substrate was hydrolysed much faster by PG 2. Both enzymes were active on pectins with different degrees of esterification, they were sensitive towards Ca-cations and not glycosylated. The kinetic properties were measured by viscosimetry with polygalacturonic acid as a substrate. NMR experiments on a model substrate revealed an inverting mechanism of carbohydrate hydrolysis for both enzymes.
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spelling Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsiiPolygalacturonaseFFEPurificationPectinasePlant pathogen fungusScience & TechnologySclerotium rolfsii (strain CBS 350.80) was found to produce extraordinary high amounts of polygalacturonases (PGs). Two of these extracellular enzymes were purified by a recently introduced preparative electrophoretic device (isoelectric focusing mode of free flow electrophoresis). PG 1 (39.5 kDa, pI 6.5) and PG 2 (38 kDa, pI 5.4) exhibited quite similar properties, they were found to be both endo-acting enzymes. Both PGs cleaved penta- and trigalacturonic acid while tetragalacturonic acid was only cleaved when trigalacturonic acid was present. The latter substrate was hydrolysed much faster by PG 2. Both enzymes were active on pectins with different degrees of esterification, they were sensitive towards Ca-cations and not glycosylated. The kinetic properties were measured by viscosimetry with polygalacturonic acid as a substrate. NMR experiments on a model substrate revealed an inverting mechanism of carbohydrate hydrolysis for both enzymes.ElsevierUniversidade do MinhoSchnitzhofer, WWeber, H.-J.Vršanská, M.Biely, P.Paulo, Artur CavacoGuebitz, G. M.2007-062007-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/13560eng0141-022910.1016/j.enzmictec.2006.11.005http://www.sciencedirect.com/science/article/pii/S0141022906005473info: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:RCAAP2023-07-21T12:45:59Zoai:repositorium.sdum.uminho.pt:1822/13560Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:43:55.840668Repositório 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 Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
title Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
spellingShingle Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
Schnitzhofer, W
Polygalacturonase
FFE
Purification
Pectinase
Plant pathogen fungus
Science & Technology
title_short Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
title_full Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
title_fullStr Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
title_full_unstemmed Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
title_sort Purification and mechanistic characterisation of two polygalacturonases from Sclerotium rolfsii
author Schnitzhofer, W
author_facet Schnitzhofer, W
Weber, H.-J.
Vršanská, M.
Biely, P.
Paulo, Artur Cavaco
Guebitz, G. M.
author_role author
author2 Weber, H.-J.
Vršanská, M.
Biely, P.
Paulo, Artur Cavaco
Guebitz, G. M.
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Schnitzhofer, W
Weber, H.-J.
Vršanská, M.
Biely, P.
Paulo, Artur Cavaco
Guebitz, G. M.
dc.subject.por.fl_str_mv Polygalacturonase
FFE
Purification
Pectinase
Plant pathogen fungus
Science & Technology
topic Polygalacturonase
FFE
Purification
Pectinase
Plant pathogen fungus
Science & Technology
description Sclerotium rolfsii (strain CBS 350.80) was found to produce extraordinary high amounts of polygalacturonases (PGs). Two of these extracellular enzymes were purified by a recently introduced preparative electrophoretic device (isoelectric focusing mode of free flow electrophoresis). PG 1 (39.5 kDa, pI 6.5) and PG 2 (38 kDa, pI 5.4) exhibited quite similar properties, they were found to be both endo-acting enzymes. Both PGs cleaved penta- and trigalacturonic acid while tetragalacturonic acid was only cleaved when trigalacturonic acid was present. The latter substrate was hydrolysed much faster by PG 2. Both enzymes were active on pectins with different degrees of esterification, they were sensitive towards Ca-cations and not glycosylated. The kinetic properties were measured by viscosimetry with polygalacturonic acid as a substrate. NMR experiments on a model substrate revealed an inverting mechanism of carbohydrate hydrolysis for both enzymes.
publishDate 2007
dc.date.none.fl_str_mv 2007-06
2007-06-01T00:00:00Z
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/1822/13560
url http://hdl.handle.net/1822/13560
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0141-0229
10.1016/j.enzmictec.2006.11.005
http://www.sciencedirect.com/science/article/pii/S0141022906005473
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 Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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