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Purification and characterization of an alpha-galactosidase from Aspergillus fumigatus BABT
Rezende,Sebastião Tavares de; Guimarães,Valéria Monteze; Rodrigues,Marília de Castro; Felix,Carlos Roberto.
Aspergillus fumigatus secreted invertase (beta-fructofuranosidase) and alpha-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). alpha-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtration and two ion exchange chromatographies and showed Mw of 54.7 kDa. The purified enzyme showed maximal activity against p-nitrophenyl-alpha-D-galactopyranoside (pNPGal) at pH 4.5-5.5 and 55 °C, and retained about 80% of the original activity after incubation for 90 minutes at 50ºC. The KM for pNPGal was 0.3 mM. Melibiose was hydrolyzed by the enzyme but raffinose was very poor substrate.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Aspergillus fumigatus; Alpha-galactosidase; Raffinose oligosaccharides.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132005000200005
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Soybean seed galactinol synthase activity as determined by a novel colorimetric assay Rev. Bras. Fisiol. Veg.
RIBEIRO,MARLUCI; FELIX,CARLOS R.; LOZZI,SILENE DE PAULINO.
Galactinol synthase (GS) is a key enzyme for the biosynthesis of raffinose oligosaccharides (RO) which are the flatulence factors present in soybean seeds and several other legumes. Understanding of soybean seed GS properties is, therefore, of biotechnological interest. The GS enzyme catalyses formation of galactinol and UDP from UDP-gal and myo-inositol. This enzyme is currently assayed by an isotopic method. We have then idealized a more convenient method for GS assay based on the indirect colorimetric determination of the UDP formed which is then hydrolyzed by exogenous apyrase and the resulting Pi quantified by a modification of the colorimetric method of Fiske & SubbaRow. The color developed is stable, and the method is suitable for detection...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Flatulence; Galactinol synthase; Colorimetric assay; Raffinose oligosaccharides.
Ano: 2000 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-31312000000300004
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