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

功能化糖核苷酸供体的​​合成，用于基于聚糖微阵列的植物糖基转移酶的鉴定和表征

• 批准号: 424354966
• 负责人: Professor Dr. Fabian Pfrengle
• 金额: --
• 依托单位: Department für Chemie (DCH)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/424354966?language=en
• 关键词: Synthesis; Functionalized; Sugar; Nucleotide; Donors

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.

植物细胞壁由一个复杂的复合材料组成，主要由几种多糖网络组成，在植物生命周期中具有重要功能。这些细胞壁多糖作为可持续材料的来源以及生产生物燃料的浓厚兴趣。为了提高利用生物量作为可再生资源的经济可行性，生成了越来越多的具有多糖多糖组成的植物。然而，执行靶向遗传修饰的先决条件是细胞壁多糖生物合成的详细知识。我们最近生产了一个配备合成细胞壁寡糖的聚糖微阵列。这种微阵列首次提供了开发各种植物糖基转移酶同时筛查的测定法。微阵列将与化学合成的氮杂型糖核苷酸和假定的糖基转移酶一起孵育。通过使用点击化学的荧光染料随后的标记，将对任何合并的氮杂官能化的单糖。因此，这种高通量测定法的微阵列格式不仅对于识别新的糖基转移酶很有价值，而且还将直接提供有关其底物特异性的信息。