Oligosaccharide structures formed during the hydrolysis of lactose by Aspergillus oryzae β-galactosidase

Di-, tri-, tetra-, penta- and hexasaccharides were formed during the hydrolysis of lactose by transgalactosylation reaction of Aspergillus oryzae β-galactosidase. In this study the isolation and characterization of the major constituents of tri-, tetra- and pentasaccharides are described. The structure elucidation of 3 tri-, 2 tetra- and 1 pentasaccharides was carried out by methylation analysis, mass spectrometry and ¹³C-nmr spectrometry. The trisaccharides are $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 3)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-}\text{d}\text{-}\text{glucose}\text{(3′-}\text{galactosyl-lactose}\text{)}$, $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-}\text{d}\text{-}\text{glucose}\text{(6′-}\text{galactosyl-lactose}\text{)}$ and $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)]-}\text{d}\text{-}\text{glucose}\text{(4,6-}\text{digalactosyl-glucose}\text{)}$. Tetrasaccharides are $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-}\text{d}\text{-}\text{glucose}$ and $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 3)}$ or $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 3)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)]-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-}\text{d}\text{-}\text{glucose}$. Pentasaccharide is $\text{O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 6)-O-β-}\text{d}\text{-}\text{galactopyranosyl}\text{-(1 → 4)-}\text{d}\text{-}\text{glucose}$.