Heterologous production and secretion of Clostridium perfringens β-toxoid in closely related Gram-positive hosts
Heterologous production and secretion of Clostridium perfringens β-toxoid in closely related Gram-positive hosts
复制标题
DOI：
10.1016/j.jbiotec.2006.07.014
复制
发表时间：
2007-01-10
影响因子：
4.1
通讯作者：
Kuipers, Oscar P.
中科院分区：
文献类型：
--
作者：
Nijland, Reindert;Lindner, Cordula;Kuipers, Oscar P.
产气荚膜梭菌是一种广泛存在的病原体。针对B和C型产气荚膜梭菌的疫苗目前使用由毒性产气荚膜梭菌菌株分泌的β-毒素来制造。从强毒株大规模生产疫苗需要严格的安全条件和昂贵的解毒和控制步骤。因此，在安全的生产宿主中以免疫原性但无毒的形式（类毒素）生产这种毒素将是有益的。为了高水平表达β-类毒素，我们在广泛宿主范围的多拷贝质粒中克隆了枯草芽孢杆菌的高活性核糖体rpsF启动子。在B。枯草杆菌，我们获得了高的细胞内生产，高达200 μ g ml（-1）的培养物。然而，β-类毒素分泌不良。所采用的rpsF表达系统允许在其他异源宿主如乳酸乳球菌和肺炎链球菌中使用相同的表达质粒。在这些生物体中，β-类毒素的分泌比最佳生产B高十倍。枯草菌这些结果显示了基于rpsF的广泛宿主范围表达系统的有用性。(c)2006 Elsevier B. V.保留所有权利。
The spore forming bacterium Clostridium perfringens is a widely occurring pathogen. Vaccines against C perfringens type B and C are currently manufactured using beta-toxin secreted by virulent C perfringens strains. Large-scale production of vaccines from virulent strains requires stringent safety conditions and costly detoxification and control steps. Therefore, it would be beneficial to produce this toxin in a safe production host and in an immunogenic, but non-toxic form (toxoid). For high-level expression of beta-toxoid, we cloned the highly active ribosomal rpsF promoter of Bacillus subtilis in a broad host range multicopy plasmid. In B. subtilis, we obtained high intracellular production, up to 200 mu g ml(-1) culture. However, the beta-toxoid was poorly secreted. The employed rpsF expression system allowed using the same expression plasmids in other heterologous hosts such as Lactococcus lactis and Streptococcus pneumoniae. In these organisms secretion of beta-toxoid was ten times higher compared to the best producing B. subtilis strain. These results show the usefulness of the rpsF based broad host range expression system. (c) 2006 Elsevier B.V. All rights reserved.