Molecular characterization of a Clostridium difficile bacteriophage and its cloned biologically active endolysin
Molecular characterization of a Clostridium difficile bacteriophage and its cloned biologically active endolysin
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DOI：
10.1128/jb.00686-08
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发表时间：
2008-10-01
影响因子：
3.2
通讯作者：
Gasson, Michael J.
中科院分区：
文献类型：
--
作者：
Mayer, Melinda J.;Narbad, Arjan;Gasson, Michael J.
艰难梭菌感染的频率和严重程度都在增加，新的高毒力菌株的出现突出了对更快速有效的控制方法的需求。在这里，我们表明，噬菌体内溶素可以用来抑制和杀死C。很难一种新的抗C.艰难梭菌，噬菌体内溶素基因的亚克隆和表达在大肠杆菌。部分纯化的内溶素对30种不同的C.艰难梭菌，并且重要的是，该组包括主要流行核糖体型027（B1/NAP 1）的菌株。相反，一系列栖息在胃肠道中的细菌物种，包括梭菌样厚壁菌门的几个代表，对内溶素不敏感。这种内溶素提供了一个平台，用于产生治疗和检测系统，以打击C。困难的问题。为了研究一种用于在胃肠道中保护性递送和生产溶素的方法，我们证明了活性CD 27 L内溶素在乳酸菌乳酸乳球菌MG 1363中的表达。
Clostridium difficile infection is increasing in both frequency and severity, with the emergence of new highly virulent strains highlighting the need for more rapid and effective methods of control. Here, we show that bacteriophage endolysin can be used to inhibit and kill C. difficile. The genome sequence of a novel bacteriophage that is active against C. difficile was determined, and the bacteriophage endolysin gene was subcloned and expressed in Escherichia coli. The partially purified endolysin was active against 30 diverse strains of C. difficile, and importantly, this group included strains of the major epidemic ribotype 027 (B1/NAP1). In contrast, a range of commensal species that inhabit the gastrointestinal tract, including several representatives of the clostridium-like Firmicutes, were insensitive to the endolysin. This endolysin provides a platform for the generation of both therapeutic and detection systems to combat the C. difficile problem. To investigate a method for the protected delivery and production of the lysin in the gastrointestinal tract, we demonstrated the expression of active CD27L endolysin in the lactic acid bacterium Lactococcus lactis MG1363.