Purificação de lacases PPO-I de Botryosphaeria rhodina

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dc.contributor.authorHiroito Obara, Francis Widmann
dc.contributor.authorVaréa-Pereira, Geni
dc.contributor.authorTomoe Miyagui, Dalva
dc.contributor.authorCorradi Da Silva, Maria de Lourdes [UNESP]
dc.contributor.institutionUniversidade Estadual de Londrina (UEL)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-27T11:21:23Z
dc.date.available2014-05-27T11:21:23Z
dc.date.issued2005-07-01
dc.description.abstractLaccases are glycoprotein polyphenol oxidases which are involved in fungal pathogenicity and they are also useful for biotechnological applications. The ligninolytic ascomycete, Botryosphaeria rhodina, has been studied as producer of exopolysaccharide and PPO-I and PPO-II laccases induced by veratryl alcohol. However, as the induced laccases have not been isolated, the aim of this study was to purify the enzyme and to identify the carbohydrates constituents of the glycosidic moiety. The fungus was cultivated on broth Vogel, 1% glucose and 30.4mM veratryl alcohol during 4.5 days at 28°C/180 rpm. The extracellular fluid showed high carbohydrate concentration and the stability of PPO-I laccase under conditions of refrigeration and freezing at 4°C-18°C over 40 days. The purification was developed by ultrafiltration using a NMWL 100 and 30 kDa membrane, gelfiltration on Sephadex G-100, and ion-exchange chromatography on DEAE-cellulose. The purified laccase was identified as a glycoprotein, weight molecular 113 kDa, consisting of 40% protein and 60% carbohydrate identified by HPAEC-PAD as fucose, galactose, mannose, glucose and glucosamine.en
dc.description.affiliationDepartamento de Bioquímica e Biotecnologia Universidade Estadual de Londrina (UEL) Campus Universitário, Cx. Postal 6001, 86051-990, Londrina, Paraná
dc.description.affiliationDepartamento de Física, Química e Biologia Faculdade de Ciências e Tecnologia Universidade Estadual Paulista (Unesp), Rua Roberto Simonsen, 305, 19060-900, Presidente Prudente, SP
dc.description.affiliationUnespDepartamento de Física, Química e Biologia Faculdade de Ciências e Tecnologia Universidade Estadual Paulista (Unesp), Rua Roberto Simonsen, 305, 19060-900, Presidente Prudente, SP
dc.format.extent303-310
dc.identifierhttp://dx.doi.org/10.4025/actascibiolsci.v27i3.1317
dc.identifier.citationActa Scientiarum - Biological Sciences, v. 27, n. 3, p. 303-310, 2005.
dc.identifier.doi10.4025/actascibiolsci.v27i3.1317
dc.identifier.file2-s2.0-33645714170.pdf
dc.identifier.issn1679-9283
dc.identifier.scopus2-s2.0-33645714170
dc.identifier.urihttp://hdl.handle.net/11449/68324
dc.language.isopor
dc.relation.ispartofActa Scientiarum: Biological Sciences
dc.relation.ispartofsjr0,208
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectBotryosphaeria rhodina
dc.subjectCharacterization
dc.subjectExopolysaccharides
dc.subjectGlycoprotein
dc.subjectLaccase
dc.subjectPurification
dc.subjectexopolysaccharide
dc.subjectfucose
dc.subjectgalactose
dc.subjectglucosamine
dc.subjectglucose
dc.subjectglycoprotein
dc.subjectlaccase
dc.subjectpolyphenol
dc.subjectsephadex
dc.subjectAscomycetes
dc.subjectbiotechnology
dc.subjectenzyme isolation
dc.subjectenzyme purification
dc.subjectenzyme stability
dc.subjectextracellular fluid
dc.subjectfungal virulence
dc.subjectfungus
dc.subjectgel filtration
dc.subjection exchange chromatography
dc.subjectnonhuman
dc.subjectultrafiltration
dc.subjectAscomycota
dc.subjectFungi
dc.titlePurificação de lacases PPO-I de Botryosphaeria rhodinapt
dc.title.alternativePurification of laccases PPO-I of fungus Botryosphaeria rhodinaen
dc.typeArtigo
dcterms.licensehttp://periodicos.uem.br/ojs/index.php/ActaSciBiolSci/about/editorialPolicies#openAccessPolicy
unesp.campusUniversidade Estadual Paulista (Unesp), Faculdade de Ciências e Tecnologia, Presidente Prudentept

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