Carbohydrate Research, 248 (1993) 285-301 Elsevier Science Publishers B.V., Amsterdam 285 Structural analysis of tamarind seed xyloglucan oligosaccharides using p-galactosidase digestion and spectroscopic methods * William S. York +, Lisa K. Harvey, Rafael Guillen, Peter Albersheim and Alan G. Darvill Complex Carbohydrate Research Center, The University of Georgia, 220 Riverbend Road, Athens GA 30602 (USA) (Received March lOth, 1993; accepted May 18th, 1993) ABSTRACT The borohydride-reduced forms (oligoglycosyl alditols) of two isomeric octasaccharides (Glc,Xyl,Gal) that are released from xyloglucans of various plant species upon treatment with a fungal endo-(1 + 4)+glucanase were isolated and structurally characterized. A mixture of oligosaccharides that is released from tamarind seed xyloglucan by the endo-(1 + 4)-/3-glucanase was digested with a commercially available P-galactosidase (Aspergillus niger). The &galactosidase selectively hydrolyzed the galactosyl residue of one of the two isomeric octasaccharides present in the mixture. A homoge- neous preparation of the p-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,Xyl,) made it possible to completely assign their ‘H and 13C NMR spectra. In addition, the borohydride-re- duced form of the p-D-galactosidase-susceptible octasaccharide isomer was purified by high pH anion-exchange chromatography of the e&o-(1 + 4)-P-glucanase-released octasaccharides from rape- seed xyloglucan (no B-galactosidase treatment), and its ‘H 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. INTRODUCTION Xyloglucans (XGs) are a class of hemicellulosic polysaccharides that are found in noncovalent association with microcrystalline cellulose fibers in the cell walls of higher plantslm4. The xyloglucan backbone consists of B-(1 + 4%linked D-Glcp * This is number XXXVI of the series entitled, The Structure of Plant Cell Walls. + Corresponding author. 0008-6215/93/$06.00 0 1993 - Elsevier Science Publishers B.V. All rights reserved