Synthesis of Saccharide Chain by Using Cells and the Conversion to Functional Polymer

为了建立生物生产大量含有GM 3型寡糖的有价值物质的最佳条件，合成了两种在不同位置具有叠氮基的乳糖苷引物，并将其导入B16黑色素瘤细胞。引物I和引物II的合成分别通过12-叠氮基-1-十二烷醇和2-叠氮基-1-十二烷醇与乳糖过乙酸酯的糖基化和随后的脱乙酰化完成。引物II（2-叠氮基十二烷基β-乳糖苷）的糖基化产物的量几乎是引物I（12-叠氮基十二烷基β-乳糖苷）的两倍。考察了接种细胞数、引物浓度和孵育时间对糖基化效率的影响。研究结果对优化叠氮法生产GM 3型低聚糖的工艺条件具有重要意义 关于我们 十二烷基乳糖苷引物和B16细胞。荧光标记的糖引物可以作为合成寡糖的可行支架。本研究通过化学方法合成了一种含氟乳糖苷引物6-（全氟己基）己基-4-O-（β-D-吡喃半乳糖基）-β-D-吡喃葡萄糖苷，并将其导入小鼠B16细胞，启动寡糖合成。B16细胞对引物的摄取导致末端半乳糖残基的唾液酸化，得到与神经节苷脂GM 3具有相同聚糖结构的寡糖。许多氟原子的存在对细胞没有任何不利影响。此外，氟原子的数量并不构成空间位阻，相反，它们的存在可能增加了引物的疏水性并增强了膜的渗透性。这种使用荧光标记的引物和细胞的策略可以为通过环境友好的方法制备寡糖铺平道路，该方法消除了大量有机溶剂的使用。少

In order to establish the optimum condition for the bio-production of a large amount of valuable materials containing GM3-type oligosaccharides, two kinds of lactoside primers having azido group at different positions were synthesized and introduced into B16 melanoma cells. The synthesis of primers I and II was accomplished by glycosylation of 12-azido-1-dodecanol and 2-azido-1-dodecanol, respectively, with lactose peracetate and subsequent deacetylation.Both primers were glycosylated to give GM3-type oligosaccharide derivatives which were released to the culture medium. The amount of glycosylated product from primer II (2-azidododecyl β-lactoside) was almost twice as compared with that from primer I (12-azidododecyl β-lactoside). The effects of seeded cell number, primer concentration and length of incubation time on the glycosylation efficiency were also investigated. The results are necessary to optimize the conditions for the mass-production of GM3-type oligosaccharide using azido … More dodecyl lactoside primer and B16 cells. Fluorous-tagged saccharide primers could be viable scaffolds for the synthesis of oligosaccharides. This research demonstrates that a fluorine-containing saccharide derivative could actually be taken up by the cell, the saccharide chain elongated by cellular enzymes, and the elongated product released by the cells to the culture medium.A fluorous-tagged lactoside primer, 6-(perfluorohexyl)hexyl-4-O-(β-D-galactopyranosyl)-β-D-glucopyranoside, was chemically synthesized and introduced in mouse B16 cells to prime oligosaccharide synthesis. Uptake of the primer by B16 cells resulted in the sialylation of the terminal galactose residue to afford an oligosaccharide with the same glycan structure as ganglioside GM3. The presence of many fluorine atoms did not have any adverse effects to the cells. Moreover, the number of fluorine atoms did not pose a steric barrier and instead, their presence possibly increased the hydrophobicity of the primer and enhanced membrane permeability. This strategy of using a fluorous-tagged primer and cells can pave the way for an easier way of preparing oligosaccharides via an environment-friendly approach that eliminates the use of large amounts of organic solvents. Less