Chemoenzymatic synthesis of bacterial polysaccharides

细菌多糖的化学酶法合成 项目摘要 细菌细胞表面装饰有显著变化的多糖结构，包括荚膜， 多糖（CPS）、O-多糖（O-PS）和胞外多糖（EPS）。这些多糖 介导细菌和宿主之间的相互作用。许多是重要的毒力因子， 介质或免疫调节剂。因为它们的组成与宿主聚糖的组成显著不同， 多种细菌多糖已被用于开发疫苗以保护人类免受多种多样的 细菌性传染病迄今为止，这些多糖通常从细菌中分离出来， 文化，因此由异质混合物组成。结构确定的多糖， 重复单元的数目不容易获得。结合三个小组的专业知识， 一锅多酶化学酶法合成寡糖重复单元，聚合 脂寡糖重复单元，用于形成Wzy依赖性细菌多糖，以及 细菌多糖、结构确定的多糖的多糖转移酶催化的生产 将合成具有指定数目的寡糖重复单元。这些化合物很有价值 用于聚糖微阵列研究的探针和用于诊断，免疫调节剂， 和疫苗。将确定最佳储存和反应条件。酶和试剂将 组装在易于储存和易于使用的套件中。合成和纯化的方案将是 通过发表论文和在指定网站上建立和共享。这些工具包将允许 非专业人士进行综合。

Chemoenzymatic synthesis of bacterial polysaccharides Project Summary The bacterial cell surface is decorated with remarkable variations of polysaccharide structures including capsular polysaccharides (CPS), O-polysaccharides (O-PS), and exopolysaccharides (EPS). These polysaccharides mediate interactions between the bacterium and its host. Many are important virulence factors, adhesion mediators, or immunomodulators. Because their composition differs dramatically from that of host glycans, multiple bacterial polysaccharides have been used to develop vaccines to protect human from a diverse array of bacterial infectious diseases. To date, these polysaccharides have been commonly isolated from the bacterial cultures and therefore consist of heterogeneous mixtures. Structurally defined polysaccharides with defined numbers of repeating units are not readily available. With the combined expertise of three groups on efficient one-pot multienzyme chemoenzymatic synthesis of oligosaccharide repeating units, polymerization of lipooligosaccharide repeating units for the formation of Wzy-dependent bacterial polysaccharides and polysaccharide synthase-catalyzed production of bacterial polysaccharides, structurally defined polysaccharides with designated numbers of oligosaccharide repeating units will be synthesized. These compounds are valuable probes for glycan microarray studies and candidates for the development of diagnostics, immunomodulators, and vaccines. Optimal storage and reaction conditions will be identified. Enzymes and reagents will be assembled in convenient-to-store and easy-to-use kits. Protocols for synthesis and purification will be established and shared by publication of papers and on a designated website. These kits will allow non-specialists to carry out the synthesis.