Structural analysis of tamarind seed xyloglucan oligosaccharides using β-galactosidase digestion and spectroscopic methods

The borohydride-reduced forms (oligoglycosyl alditols) of two isomeric octasaccharides (Glc 4Xyl 3Gal) that are released from xyloglucans of various plant species upon treatment with a fungal endo-(1 → 4)-β-glucanase were isolated and structurally characterized. A mixture of oligosaccharides that is released from tamarind seed xyloglucan by the endo-(1 → 4)-β-glucanase was digested with a commercially available β-galactosidase ( Aspergillus niger). The β-galactosidase selectively hydrolyzed the galactosyl residue of one of the two isomeric octasaccharides present in the mixture. A homogeneous preparation of the β-galactosidase-resistant octasaccharide was prepared by high-resolution gel-permeation chromatography of the enzyme-digestion products. Spectroscopic characterization of the oligoglycosyl alditol prepared by reduction of this octasaccharide confirmed the previously proposed structure that had been based on analysis of the mixture of isomeric octasaccharides. The availability of large amounts of the pure, reduced octasaccharide and of a pure, reduced pentasaccharide (Glc 3Xyl 2) made it possible to completely assign their 1H and 13C NMR spectra. In addition, the borohydride-reduced form of the β- d-galactosidase-susceptible octasaccharide isomer was purified by high pH anion-exchange chromatography of the endo-(1 → 4)-β-glucanase-released octasaccharides from rapeseed xyloglucan (no β-galactosidase treatment), and its 1H and 13C NMR spectra were assigned. Additional correlations between specific structural features of xyloglucan oligoglycosyl alditols and the positions of specific resonances in their NMR spectra were deduced and added to the extensive list that we have compiled. The effects of recording the NMR spectra of the xyloglucan oligoglycosyl alditols in the presence of borate salts, which could lead to incorrect structural assignments, are also described.