Whole cell- and protein-based biosensors for the detection of bioavailable heavy metals in environmental samples
Whole cell- and protein-based biosensors for the detection of bioavailable heavy metals in environmental samples
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DOI：
10.1016/s0003-2670(98)00725-9
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发表时间：
1999-04-28
影响因子：
6.2
通讯作者：
Mattiasson, B
中科院分区：
文献类型：
--
作者：
Corbisier, P;van der Lelie, D;Mattiasson, B
这项工作的主要目标是建立两种具有增强特异性和选择性的生物传感器技术的可行性，用于检测环境样品中的几种生物可利用的重金属。遵循了两种并行策略。第一种方法是构建全细胞细菌生物传感器，在存在生物可用重金属的情况下发出生物发光或荧光信号。 sigma-54启动子作为环境污染物传感元件的分子基础已经确定，并且许多金属诱导的启动子区域已经被鉴定、测序并克隆为启动子盒。启动子盒的特异性已使用 luxCDABE 报告系统确定。含有金属诱导勒克斯报告系统的全细胞生物传感器已被纳入不同的基质中，以便随后固定在光纤上，并根据其灵敏度和存储容量进行表征。第二种类型的审查基于金属结合蛋白和重金属离子之间的直接相互作用。在这种情况下，在飞摩尔至毫摩尔金属离子浓度存在的情况下，检测到聚球藻金属硫蛋白（作为 GST-SmtA 融合蛋白）和汞调节蛋白 MerR 等蛋白质的电容变化。 (C) 1999 Elsevier Science B.V. 保留所有权利。
The principal goal of this work was to establish the feasibility of two biosensor technologies with enhanced specificity and selectivity for the detection of several bioavailable heavy metals in environmental samples. Two parallel strategies have been followed. The first approach was to construct whole cell bacterial biosensors that emit a bioluminescent or fluorescent signal in the presence of a biologically available heavy metal. The molecular basis of sigma-54 promoters as sensing elements of environmental pollutants has been determined and a number of metal-induced promoter regions have been identified, sequenced and cloned as promoter cassettes. The specificity of the promoter cassettes has been determined using luxCDABE reporter systems. Whole cell-biosensors containing metal-induced lux reporter systems have been incorporated into different matrices for their later immobilisation on optic fibres and characterised in terms of their sensitivity and storage capacity. The second type of censors was based on the direct interaction between metal-binding proteins and heavy metal ions. In this case, the capacitance changes of the proteins, such as synechoccocal metallothionein (as a GST-SmtA fusion protein) and the mercury regulatory protein, MerR, were detected in the presence of femtomolar to millimolar metal ion concentrations. (C) 1999 Elsevier Science B.V. All rights reserved.