Título: | Purification and characterization of soy cotyledon B-glucosidase. |
Autores: |
SANTOS, R. F. OLIVEIRA, C. F. VARÉA, G. S. SILVA, M. L. C. ORRADI da IDA, E. I. MANDARINO, J. M. G. CARRÃO-PANIZZI, M. C. RIBEIRO, M. L. L. |
Fecha: |
2013 2012-07-10 |
Publicador: | Repositório Acesso Livre à Informação Científica da Embrapa (ALICE) |
Fuente: | |
Tipo: | Artigo em periódico indexado (ALICE) |
Tema: | Soybean |
Descripción: |
b-Glucosidase F42 of soy cotyledons was purified by ammonium sulfate fractionation, ion-exchange chromatography (CM-Sephadex-C-50, Sigma, St. Louis, MO) and gel filtration (Sephadex G-100, Sigma). The enzyme was purified 111.8-fold relative to its concentration in the crude extract. It had an apparent molecular mass of 53 kDa in gel filtration experiments and produced a 33-kDa band in sodium dodecyl sulfate?polyacrylamide gel electrophoresis, suggesting that it is dimeric. The purified b-glucosidase F42 was characterized as a glycoprotein after the identification of fucose, galactosamine and glucosamine by high-pressure anion-exchange chromatography?pulsed amperometric detector. Its highest activity was observed at pH 5.0 and 45C, and it was stable for up to 4 days at 25C. The Km of the enzyme was 0.12 mMp-nitrophenyl-b-d-glucopyranoside. b-Glucosidase F42 showed specificity for different substrates, and its activity was inhibited by 1 mM HgCl2, 10mM glucono-d-lactone or 150 mMglucose and increased by 10 mMMnCl2. 2013 |
Idioma: | en |