Synthesis of Functionalized Sugar Nucleotide Donors for the Glycan-Microarray-Based Identification and Characterization of Plant Glycosyltransferases

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Plant cell walls consist of a sophisticated composite largely made of several polysaccharide networks with essential functions in the life cycle of the plant. These cell wall polysaccharides receive an enormous interest as sources of sustainable materials and for the production of biofuels. To enhance the economic viability of exploiting biomass as a renewable resource, an increasing number of plants with modified polysaccharide composition are generated. However, a prerequisite to perform targeted genetic modifications is a detailed knowledge of cell wall polysaccharide biosynthesis. We recently produced a glycan microarray equipped with synthetic cell wall oligosaccharides. This microarray provides for the first time the opportunity to develop an assay for the simultaneous screening of various plant glycosyltransferases. The microarray will be incubated with chemically synthesized azido-functionalized sugar nucleotides and putative glycosyltransferases. Any incorporated azido-functionalized monosaccharide will be visualized by subsequent labeling with a fluorescent dye using click-chemistry. Thus, the microarray format of this high-throughput assay will not only be valuable for identifying new glycosyltransferases, but will directly provide information on their substrate specificities.