Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, UK.
Microb Biotechnol. 2011 Jan;4(1):89-97. doi: 10.1111/j.1751-7915.2010.00228.x.
We developed a biocompatible and highly efficient approach for functionalization of bacterial cell wall with magnetic nanoparticles (MNPs). Three Acinetobacter baylyi ADP1 chromosomally based bioreporters, which were genetically engineered to express bioluminescence in response to salicylate, toluene/ xylene and alkanes, were functionalized with 18 3 nm iron oxide MNPs to acquire magnetic function. The efficiency of MNPs functionalization of Acinetobacter bioreporters was 99.96 0.01%. The MNPs-functionalized bioreporters (MFBs) can be remotely controlled and collected by an external magnetic field. The MFBs were all viable and functional as good as the native cells in terms of sensitivity, specificity and quantitative response. More importantly, we demonstrated that salicylate sensing MFBs can be applied to sediments and garden soils, and semiquantitatively detect salicylate in those samples by discriminably recovering MFBs with a permanent magnet. The magnetically functionalized cells are especially useful to complex environments in which the indigenous cells, particles and impurities may interfere with direct measurement of bioreporter cells and conventional filtration is not applicable to distinguish and harvest bioreporters. The approach described here provides a powerful tool to remotely control and selectively manipulate MNPs-unctionalized cells in water and soils. It would have a potential in the application of environmental microbiology, such as bioremediation enhancement and environment monitoring and assessment.
我们开发了一种生物相容性的高效方法,用于将磁性纳米颗粒 (MNPs) 功能化细菌细胞壁。三种基于 Acinetobacter baylyi ADP1 染色体的生物报告基因,通过基因工程设计,可对水杨酸、甲苯/二甲苯和烷烃作出生物发光响应,被功能化的 18nm 氧化铁 MNPs 赋予了磁性功能。Acinetobacter 生物报告基因的 MNPs 功能化效率为 99.96 0.01%。MNPs 功能化的生物报告基因(MFBs)可以通过外部磁场进行远程控制和收集。在灵敏度、特异性和定量响应方面,MFBs 与原生细胞一样具有良好的活性和功能。更重要的是,我们证明了水杨酸感应 MFBs 可应用于沉积物和花园土壤,并通过使用永久磁铁可辨别性地回收 MFBs 来对半定量检测这些样品中的水杨酸。磁性功能化细胞对于复杂环境尤其有用,在这些环境中,土著细胞、颗粒和杂质可能会干扰生物报告基因细胞的直接测量,并且常规过滤不适用于区分和收获生物报告基因细胞。这里描述的方法为在水和土壤中远程控制和选择性操作 MNPs 功能化细胞提供了一种强大的工具。它在环境微生物学中的应用具有潜力,例如生物修复增强和环境监测和评估。