AI-2 Biosynthesis Module in a Magnetic Nanofactory Alters Bacterial Response Via Localized Synthesis and Delivery
AI-2 Biosynthesis Module in a Magnetic Nanofactory Alters Bacterial Response Via Localized Synthesis and Delivery
复制标题
DOI：
10.1002/bit.22078
复制
发表时间：
2009-02-01
影响因子：
3.8
通讯作者：
Bentley, William E.
中科院分区：
文献类型：
--
作者：
Fernandes, Rohan;Bentley, William E.
纳米工厂是纳米尺寸的，由提供各种功能的模块组成，这些模块通过局部合成并将货物递送到目标细胞表面来改变目标细胞的反应。在其基本形式中，纳米工厂至少由两个功能模块组成：细胞捕获模块和合成模块。在这项工作中，展示了磁性纳米工厂，通过局部合成和传递“通用”细菌群体感应信号分子自诱导剂 AI-2 来改变目标细菌的反应。磁性纳米工厂由细胞捕获模块（壳聚糖-mag 纳米颗粒）和 AI-2 生物合成模块组成，该模块包含组装在一起的融合蛋白（His-LuxS-Pfs-Tyr，HLPT）上的 AI-2 生物合成酶 Pfs 和 LLIXS。由于融合蛋白内两种酶的接近性，假设 HUT 在将底物 SAH 转化为产物 AI-2 方面比其组成酶（单独使用）更有效。在广泛的实验条件下，HLPT 被证明比组成酶 Pfs 和 LuxS 更活跃。磁性纳米工厂（含有结合的 HLPT）也被证明比游离的、未结合的 HLPT 更活跃。与直接向细胞添加 AI-2 产生的反应相比，由于 AI-2 的局部合成和递送，它们还可以在目标大肠杆菌细胞中引发增强的反应。预计将开展研究调查 AI-2 的普遍性，并利用磁性纳米工厂解开细菌中基于 AI-2 的群体感应。讨论了使用这种多模块纳米工厂开发基于拦截和中断群体感应信号的下一代抗菌药物的前景。
Nanofactories are nano-dimensioned and comprised of modules serving various functions that alter the response of targeted cells when deployed by locally synthesizing and delivering cargo to the surfaces of the targeted cells. In its basic form, a nanofactory consists of a minimum of two functional modules: a cell capture module and a synthesis module. In this work, magnetic nanofactories that alter the response of targeted bacteria by the localized synthesis and delivery of the "universal" bacterial quorum sensing signal molecule autoinducer AI-2 are demonstrated. The magnetic nanofactories consist of a cell capture module (chitosan-mag nanoparticles) and an AI-2 biosynthesis module that contains both AI-2 biosynthetic enzymes Pfs and LLIXS on a fusion protein (His-LuxS-Pfs-Tyr, HLPT) assembled together. HUT is hypothesized to be more efficient than its constituent enzymes (used separately) at conversion of the substrate SAH to product AI-2 on account of the proximity of the two enzymes within the fusion protein. HLPT is demonstrated to be more active than the constituent enzymes, Pfs and LuxS, over a wide range of experimental conditions. The magnetic nanofactories (containing bound HLPT) are also demonstrated to be more active than free, unbound HLPT. They are also shown to elicit an increased response in targeted Escherichia coli cells, due to the localized synthesis and delivery of AI-2, when compared to the response produced by the addition of AI-2 directly to the cells. Studies investigating the univvrsality of AI-2 and unraveling AI-2 based quorum sensing in bacteria using magnetic nanofactories are envisioned. The prospects of using such multi-modular nanofactories in developing the next generation of antimicrobials based on intercepting and interrupting quorum sensing based signaling are discussed.