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

利用细胞合成糖链并转化为功能聚合物

基本信息

批准号：
14350483
负责人：
HATANAKA Kenichi
金额：
$ 8.13万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

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

为了建立大量含有GM3型寡糖的有价值物质的生物生产的最佳条件，合成了两种不同位置上具有叠氮基的乳糖苷引物，并将其引入B16黑色素瘤细胞中。引物I和II的合成是通过分别用全乙酸乳糖糖基化12-叠氮基-1-十二醇和2-叠氮基-1-十二醇并随后脱乙酰化来完成的。两种引物都被糖基化以得到GM3型寡糖衍生物，其被释放到培养基中。引物II（2-叠氮十二烷基β-乳糖苷）的糖基化产物的量几乎是引物I（12-叠氮十二烷基β-乳糖苷）的糖基化产物的两倍。还研究了接种细胞数、引物浓度和孵育时间长度对糖基化效率的影响。这些结果对于优化使用叠氮十二烷基乳糖苷引物和 B16 细胞大规模生产 GM3 型寡糖的条件是必要的。氟标记的糖引物可能是合成寡糖的可行支架。这项研究表明，含氟糖衍生物实际上可以被细胞吸收，糖链被细胞酶延长，延长的产物由细胞释放到培养基中。含氟标记的乳糖苷引物 6-(全氟己基)己基-4-O-(β-D-吡喃半乳糖基)-β-D-吡喃葡萄糖苷化学合成并引入小鼠 B16 细胞以启动寡糖合成。 B16 细胞摄取引物导致末端半乳糖残基唾液酸化，得到与神经节苷脂 GM3 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屏障，相反，它们的存在可能增加底漆的疏水性并增强膜的渗透性。这种使用氟标记引物和细胞的策略可以为通过环境友好的方法制备低聚糖铺平道路，从而消除大量有机溶剂的使用。较少的