Chemoenzymatic synthesis of bacterial polysaccharides

细菌多糖的化学酶法合成

基本信息

批准号：
9981827
负责人：
Laura L Kiessling
金额：
$ 72.04万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9981827
关键词：
ATP-Binding Cassette Transporters Address Adhesions Anabolism Bacteria Bacterial Polysaccharides Biological Biomedical Research Cell surface Chondroitin Communicable Diseases Complex Development Diagnosis Diagnostic Diphosphates Enzymes Galactans Generations Genus Mycobacterium Glycobiology High Pressure Liquid Chromatography Human Immune system Immunomodulators In Vitro Individual Laboratories Length Lipids Mediating Mediator of activation protein Monosaccharides Oligosaccharides Paper Pathogenicity Pathway interactions Polymerase Polysaccharides Preparation Production Protocols documentation Publications Reaction Reagent Research Research Personnel Structure System Technology Vaccine Design Vaccines Validation Variant Virulence Factors base carbohydrate binding protein chemical synthesis experience lipooligosaccharide medical specialties microbial polymerization programs reconstitution research and development web site

项目摘要

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.

细菌多糖的化学酶合成项目摘要细菌细胞表面装饰有多种多糖结构的显着变化多糖（CPS），O-多糖（O-PS）和外多糖（EPS）。这些多糖介导细菌与其宿主之间的相互作用。许多是重要的毒力因素，粘附介质或免疫调节剂。因为它们的成分与宿主聚糖的组成截然不同，所以多种细菌多糖已用于开发疫苗，以保护人免受多种多样的阵列细菌传染病。迄今为止，这些多糖通常是从细菌中分离出来的培养物，因此由异质混合物组成。结构定义的多糖具有定义重复单元的数量不容易获得。具有三组的综合知识一锅多酶化学酶合成的寡糖重复单元，聚合的聚合脂肪糖重复单元，用于形成WZY依赖性细菌多糖和多糖合成酶催化的细菌多糖的产生，结构定义的多糖将合成指定数量的寡糖重复单元。这些化合物很有价值聚糖微阵列研究和候选诊断，免疫调节剂的探针，和疫苗。将确定最佳的存储和反应条件。酶和试剂将是配备方便且易于使用的套件。合成和纯化的协议将是通过论文和指定网站上的出版物建立和共享。这些套件将允许非专家进行综合。