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Extracellular α-Galactosidase from Debaryomyces hansenii UFV-1 and Its Use in the Hydrolysis of Raffinose Oligosaccharides

Pollyanna A. Viana,^† Sebastião T. de Rezende,^† Virgínia M. Marques,^† Larissa M. Trevizano,^† Flávia M. L. Passos,^† Maria G. A. Oliveira,^† Marcelo P. Bemquerer,^‡ Jamil S. Oliveira,^‡ and Valéria M. Guimarães*^†

BIOAGRO, Universidade Federal de Viosa, Viosa, MG, 36570-000, and Departamento de Bioqumica e Imunologia, Universidade Federal de Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil

J. Agric. Food Chem., 2006, 54 (6), pp 2385–2391

DOI: 10.1021/jf0526442

Publication Date (Web): February 17, 2006

†

Universidade Federal de Viçosa.

‡

Universidade Federal de Minas Gerais.

To whom correspondence should be addressed. Tel: (5531)3899-2374. Fax: (5531)3899-2373. E-mail: vmonteze@ufv.br.

Abstract

Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestion of soybean-derived products. ROs are hydrolyzed by α-galactosidases that cleave α-1,6-linkages of α-galactoside residues. The objectives of this study were the purification and characterization of extracellular α-galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtration and anion exchange chromatographies presented an M_r value of 60 kDa and the N-terminal amino acid sequence YENGLNLVPQMGWN. The K_m values for hydrolysis of pNPαGal, melibiose, stachyose, and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The α-galactosidase presented absolute specificity for galactose in the α-position, hydrolyzing pNPGal, stachyose, raffinose, melibiose, and polymers. The enzyme was noncompetitively inhibited by galactose (K_i = 2.7 mM) and melibiose (K_i = 1.2 mM). Enzyme treatments of soy milk for 4 h at 60 °C reduced the amounts of stachyose and raffinose by 100%.

Keywords: α-Galactosidase; Debaryomyces hansenii UFV-1; raffinose oligosaccharides; characterization; flatulence

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Published In Issue March 22, 2006
Received for review October 25, 2005. Revised manuscript received January 5, 2006. Accepted January 26, 2006. This study was supported by grants of the Fundação de Amparo à Pesquisa do Estado de Minas GeraisFAPEMIG (EDT24000, EDT268/05) and Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorCAPES, Brazil.