Extracellular -Galactosidase from Debaryomyces hansenii UFV-1 and Its Use in the Hydrolysis of Raffinose Oligosaccharides
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- 作者：Pollyanna A. Viana ; Sebastiã ; o T. de Rezende ; Virgí ; nia M. Marques ; Larissa M. Trevizano ; Flá ; via M. L. Passos ; Maria G. A. Oliveira ; Marcelo P. Bemquerer ; Jamil S. Oliveira ; and Val
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2006
- 出版时间：March 22, 2006
- 年：2006
- 卷：54
- 期：6
- 页码：2385 - 2391
- 全文大小：179K
- 年卷期：v.54,no.6(March 22, 2006)
- ISSN：1520-5118
文摘
Raffinose oligosaccharides (RO) are the factors primarily responsible for flatulence upon ingestionof soybean-derived products. ROs are hydrolyzed by 
-galactosidases that cleave -1,6-linkages of-galactoside residues. The objectives of this study were the purification and characterization ofextracellular -galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtrationand anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal aminoacid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNPGal, melibiose, stachyose,and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The -galactosidase presented absolutespecificity for galactose in the -position, hydrolyzing pNPGal, stachyose, raffinose, melibiose, andpolymers. 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 
C reduced the amounts of stachyose andraffinose by 100%.Keywords: -Galactosidase; Debaryomyces hansenii UFV-1; raffinose oligosaccharides; characterization; flatulence
-galactosidases that cleave -1,6-linkages of-galactoside residues. The objectives of this study were the purification and characterization ofextracellular -galactosidase from Debaryomyces hansenii UFV-1. The enzyme purified by gel filtrationand anion exchange chromatographies presented an Mr value of 60 kDa and the N-terminal aminoacid sequence YENGLNLVPQMGWN. The Km values for hydrolysis of pNPGal, melibiose, stachyose,and raffinose were 0.30, 2.01, 9.66, and 16 mM, respectively. The -galactosidase presented absolutespecificity for galactose in the -position, hydrolyzing pNPGal, stachyose, raffinose, melibiose, andpolymers. 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 C reduced the amounts of stachyose andraffinose by 100%.Keywords: -Galactosidase; Debaryomyces hansenii UFV-1; raffinose oligosaccharides; characterization; flatulence