Characterization of the Oligosaccharides from Lipid-Linked Oligosaccharides of Mung Bean Seedlings

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Abstract
Lipid-linked oligosaccharides were synthesized with the particulate enzyme preparation from mung bean (P. aureus) seedlings in the presence of GDP-[14C] mannose. The oligosaccharides were released from the lipids by mild acid hydrolysis and purified by several passages on Biogel P-4 columns. Five different oligosaccharides were purified in this way. Based on their relative elution constants (Kd) compared to variety of standard oligosaccharides, they were sized as (mannose-acetylglucosamine) Man7GlcNAc2, Man5GlcNAc2, Man3GlcNAc2, Man2GlcNAc2, and ManGlcNAc2. These oligosaccharides were treated with endoglucosaminidase H and .alpha.- and .beta.-mannosidase and the products were examined on Biogel P-4 columns. They were subjected to a number of chemical treatments including analysis of the reducing sugar by NaB3H4 reduction, methylation analysis and, in some cases, acetolysis. The likely structures of these oligosaccharides were determined. The synthesis of the Man7GlcNAc2 was greatly diminished when tunicamycin (10 .mu.g/ml) was added to the incubation mixtures.