Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli.
Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli.
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DOI：
10.1098/rsob.150243
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发表时间：
2016-04
期刊：
影响因子：
5.8
通讯作者：
Wren BW
中科院分区：
文献类型：
--
作者：
Kay EJ;Yates LE;Terra VS;Cuccui J;Wren BW
目前，肺炎链球菌每年在全世界造成超过1400万例肺炎病例，以及超过100万例死亡，其中大多数是儿童。致病机制的主要决定因素是可变的多糖荚膜，用于根据血清型区分菌株。该胶囊构成了肺炎球菌多糖疫苗（PPV 23）和肺炎球菌结合疫苗（PCV 13）的基础，前者含有来自23种血清型的纯化荚膜多糖，后者含有与CRM 197（突变白喉毒素）结合的13种常见血清型。从S.肺炎球菌结合疫苗的生产需要肺炎球菌，成本可能高得令人望而却步，限制了低收入国家疫苗的可及性。在这项研究中，我们证明了来自四种不同血清型的S。大肠杆菌内的肺炎。此外，我们试图确定可靠和有效地表达这些胶囊异源所需的基因的最小集合。这些E.大肠杆菌菌株可用于生产供应的S.肺炎克雷伯菌型特异性胶囊，而无需培养致病菌。此外，这些菌株可以应用于合成糖生物学应用：使用E. coli外膜囊泡或使用蛋白聚糖偶联技术偶联至蛋白质。
Currently, Streptococcus pneumoniae is responsible for over 14 million cases of pneumonia worldwide annually, and over 1 million deaths, the majority of them children. The major determinant for pathogenesis is a polysaccharide capsule that is variable and is used to distinguish strains based on their serotype. The capsule forms the basis of the pneumococcal polysaccharide vaccine (PPV23) that contains purified capsular polysaccharide from 23 serotypes, and the pneumococcal conjugate vaccine (PCV13), containing 13 common serotypes conjugated to CRM197 (mutant diphtheria toxin). Purified capsule from S. pneumoniae is required for pneumococcal conjugate vaccine production, and costs can be prohibitively high, limiting accessibility of the vaccine in low-income countries. In this study, we demonstrate the recombinant expression of the capsule-encoding locus from four different serotypes of S. pneumoniae within Escherichia coli. Furthermore, we attempt to identify the minimum set of genes necessary to reliably and efficiently express these capsules heterologously. These E. coli strains could be used to produce a supply of S. pneumoniae serotype-specific capsules without the need to culture pathogenic bacteria. Additionally, these strains could be applied to synthetic glycobiological applications: recombinant vaccine production using E. coli outer membrane vesicles or coupling to proteins using protein glycan coupling technology.