Artificial Golgi Apparatus: Globular Protein-like Dendrimer Facilitates Fully Automated Enzymatic Glycan Synthesis
Artificial Golgi Apparatus: Globular Protein-like Dendrimer Facilitates Fully Automated Enzymatic Glycan Synthesis
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DOI：
10.1021/ja106955j
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发表时间：
2010-11-24
影响因子：
15
通讯作者：
Nishimura, Shin-Ichiro
中科院分区：
文献类型：
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作者：
Matsushita, Takahiko;Nagashima, Izuru;Nishimura, Shin-Ichiro
尽管合成聚糖作为生物学研究和药物发现的工具越来越重要，但缺乏常规合成的通用方法仍然是一个主要障碍。我们已经开发了一种新的自动合成聚糖的方法，采用酶的方法和树状大分子作为化学过程中的理想支持。使用中空纤维超滤模块的回收测试表明，单分散的G6 （MW = 58 kDa）和G7 （MW = 116 kDa）聚（胺胺）树状大分子表现出与BSA （MW = 66 kDa）相似的特征。G7树状大分子在水中具有高溶解度和低粘度的特点，大大提高了自动化合成的吞吐量和效率，而基于聚丙烯酰胺的随机载体在重复反应/分离步骤中会造成重大损失。目前的方案允许在4天内以16%的总收率从简单的n -乙酰- d -葡萄糖胺连接到氨基功能化的G7树状大分子的全自动酶合成sialyl Lewis X四糖衍生物。
Despite the growing importance of synthetic glycans as tools for biological studies and drug discovery, a lack of common methods for the routine synthesis remains a major obstacle. We have developed a new method for automated glycan synthesis that employs the enzymatic approach and a dendrimer as an ideal support within the chemical process. Recovery tests using a hollow fiber ultrafiltration module have revealed that monodisperse G6 (MW = 58 kDa) and G7 (MW = 116 kDa) poly(amidoamine) dendrimers exhibit a similar profile to BSA (MW = 66 kDa). Characteristics of the globular protein-like G7 dendrimer with high solubility and low viscosity in water greatly enhanced throughput and efficiency in automated synthesis while random polyacrylamide-based supports entail significant loss during the repetitive reaction/separation step. The present protocol allowed for the fully automated enzymatic synthesis of sialyl Lewis X tetrasaccharide derivatives over a period of 4 days in 16% overall yield from a simple N-acetyl-D-glucosamine linked to an aminooxy-functionalized G7 dendrimer.