Purification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lin.
dc.contributor.author | Barata, Ricardo Andrade | |
dc.contributor.author | Andrade, Milton Hércules Guerra de | |
dc.contributor.author | Rodrigues, Roberta Dias | |
dc.contributor.author | Castro, Ieso de Miranda | |
dc.date.accessioned | 2017-10-10T14:58:42Z | |
dc.date.available | 2017-10-10T14:58:42Z | |
dc.date.issued | 2002 | |
dc.description.abstract | An 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. | pt_BR |
dc.identifier.citation | BARATA, R. A. et al. Purification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lin. Journal of Bioscience and Bioengineering USA, v. 94, n. 4, p. 304-308, 2002. Disponível em: <http://www.sciencedirect.com/science/article/pii/S1389172302801682>. Acesso: 10 jan. 2017. | pt_BR |
dc.identifier.doi | https://doi.org/10.1016/S1389-1723(02)80168-2 | |
dc.identifier.issn | 1389-1723 | |
dc.identifier.uri | http://www.repositorio.ufop.br/handle/123456789/8915 | |
dc.identifier.uri2 | http://www.sciencedirect.com/science/article/pii/S1389172302801682 | pt_BR |
dc.language.iso | en_US | pt_BR |
dc.rights | aberto | pt_BR |
dc.rights.license | O periódico Journal of Bioscience and Bioengineering concede permissão para depósito deste artigo no Repositório Institucional da UFOP. Número da licença: 3266010491564. | pt_BR |
dc.subject | Trypsin-like protease | pt_BR |
dc.subject | Fusarium | pt_BR |
dc.subject | Inhibitor effects | pt_BR |
dc.title | Purification and characterization of an extracellular trypsin-like protease of Fusarium oxysporum var. lin. | pt_BR |
dc.type | Artigo publicado em periodico | pt_BR |
Arquivos
Pacote original
1 - 1 de 1
Nenhuma Miniatura Disponível
- Nome:
- ARTIGO_PurificationCharacterizationExtracellular.pdf
- Tamanho:
- 590.37 KB
- Formato:
- Adobe Portable Document Format
Licença do pacote
1 - 1 de 1
Nenhuma Miniatura Disponível
- Nome:
- license.txt
- Tamanho:
- 924 B
- Formato:
- Item-specific license agreed upon to submission
- Descrição: