Purification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lin.

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MetadadosDescriçãoIdioma
Autor(es): dc.creatorBarata, Ricardo Andrade-
Autor(es): dc.creatorAndrade, Milton Hércules Guerra de-
Autor(es): dc.creatorRodrigues, Roberta Dias-
Autor(es): dc.creatorCastro, Ieso de Miranda-
Data de aceite: dc.date.accessioned2025-08-21T15:16:13Z-
Data de disponibilização: dc.date.available2025-08-21T15:16:13Z-
Data de envio: dc.date.issued2017-10-10-
Data de envio: dc.date.issued2017-10-10-
Data de envio: dc.date.issued2002-
Fonte completa do material: dc.identifierhttp://www.repositorio.ufop.br/handle/123456789/8915-
Fonte completa do material: dc.identifierhttp://www.sciencedirect.com/science/article/pii/S1389172302801682-
Fonte completa do material: dc.identifierhttps://doi.org/10.1016/S1389-1723(02)80168-2-
Fonte: dc.identifier.urihttp://educapes.capes.gov.br/handle/capes/1008623-
Descrição: dc.descriptionAn alkaline serineprotease, capable of hydrolyzing Nu-benzoyl-DL arginine p-nitroanilide, was secreted by Fusurium oxysporum var. hi grown in the presence of gelatin as the sole nitrogen and carbon source. The protease was purified 65-fold to electrophoretic homogenity from the culture supernatant in a three-step procedure comprising QSepharose chromatography, aMnity chromatography, and FPLC on a MonoQ column. SDS-PAGE analysis of the purified protein indicated an estimated molecular mass of 41 kDa. The protease had optimum activity at a reaction temperature of 45OC and showed a rapid decrease of activity at 48OC. The optimum pH was around 8.0. Characterization of the protease showed that Ca*+ and MgZ+ cations increased the activity, which was not inhibited by EDTA or l,lO-phenanthroline. The enzyme activity on Nubenzoyl-DL arginine p-nitroanilide was inhibited by 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride,p-aminobenzamidine dihydrochloride, aprotinin, 3-4 dichloroisocoumarin, and IVtosyl-L-lysine chloromethyl ketone. The enzyme is also inhibited by substrate concentrations higher than 2.5x lo-4 M. The protease had a Michaelis-Menten constant of 0.16 mM and a V,, of 0.60 pm01 released product .min-‘.mg’ enzyme when assayed in a non-inhibiting substrate concentration. The activity on Nu-benzoyl-DL arginine p-nitroanilide was competitively inhibited by p-aminobenzamidine dihydrochoride. A Ki value of 0.04 mM was obtained.-
Formato: dc.formatapplication/pdf-
Idioma: dc.languageen-
Direitos: dc.rightsaberto-
Direitos: dc.rightsO periódico Journal of Bioscience and Bioengineering concede permissão para depósito deste artigo no Repositório Institucional da UFOP. Número da licença: 3266010491564.-
Palavras-chave: dc.subjectTrypsin-like protease-
Palavras-chave: dc.subjectFusarium-
Palavras-chave: dc.subjectInhibitor effects-
Título: dc.titlePurification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lin.-
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