2006•Journal of Agricultural and Food ChemistryRequires access
Extracellular α-Galactosidase fromDebaryomyces hanseniiUFV-1 and Its Use in the Hydrolysis of Raffinose Oligosaccharides
Pollyanna Amaral Viana, Sebastião Tavares de Rezende, Virgínia M. Marques, Larissa Mattos Trevizano, Flávia Maria Lopes Passos, M. G. A. Oliveira, Marcelo P. Bemquerer, Jamil Silvano de Oliveira, Valéria Monteze Guimarães
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Abstract
Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestion of soybean-derived products. ROs are hydrolyzed by alpha-galactosidases that cleave alpha-1,6-linkages of alpha-galactoside residues. The objectives of this study were the purification and characterization of extracellular alpha-galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtration and anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal amino acid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNP alphaGal, melibiose, stachyose, and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The alpha-galactosidase presented absolute specificity for galactose in the alpha-position, hydrolyzing pNPGal, stachyose, raffinose, melibiose, and polymers. The enzyme was noncompetitively inhibited by galactose (Ki = 2.7 mM) and melibiose (Ki = 1.2 mM). Enzyme treatments of soy milk for 4 h at 60 degrees C reduced the amounts of stachyose and raffinose by 100%.