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Biosens Bioelectron. 2017 Aug 15;94:380-387. doi: 10.1016/j.bios.2017.03.029. Epub 2017 Mar 16.

Functional characterization of Gram-negative bacteria from different genera as multiplex cadmium biosensors.

Author information

1
Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Plenty Road, Melbourne, Victoria 3086, Australia. Electronic address: ltbereza-malcolm@students.latrobe.edu.au.
2
Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Plenty Road, Melbourne, Victoria 3086, Australia. Electronic address: sa.aracic@gmail.com.
3
Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Plenty Road, Melbourne, Victoria 3086, Australia. Electronic address: ruban_andrew@hotmail.com.
4
Land Division, Defence Science and Technology Group, Melbourne, Victoria 3207, Australia. Electronic address: gulaymann@gmail.com.
5
Applied and Environmental Microbiology Laboratory, Department of Physiology, Anatomy and Microbiology, La Trobe University, Plenty Road, Melbourne, Victoria 3086, Australia. Electronic address: A.Franks@latrobe.edu.au.

Abstract

Widespread presence of cadmium in soil and water systems is a consequence of industrial and agricultural processes. Subsequent accumulation of cadmium in food and drinking water can result in accidental consumption of dangerous concentrations. As such, cadmium environmental contamination poses a significant threat to human health. Development of microbial biosensors, as a novel alternative method for in situ cadmium detection, may reduce human exposure by complementing traditional analytical methods. In this study, a multiplex cadmium biosensing construct was assembled by cloning a single-output cadmium biosensor element, cadRgfp, and a constitutively expressed mrfp1 onto a broad-host range vector. Incorporation of the duplex fluorescent output [green and red fluorescence proteins] allowed measurement of biosensor functionality and viability. The biosensor construct was tested in several Gram-negative bacteria including Pseudomonas, Shewanella and Enterobacter. The multiplex cadmium biosensors were responsive to cadmium concentrations ranging from 0.01 to 10µgml-1, as well as several other heavy metals, including arsenic, mercury and lead at similar concentrations. The biosensors were also responsive within 20-40min following exposure to 3µgml-1 cadmium. This study highlights the importance of testing biosensor constructs, developed using synthetic biology principles, in different bacterial genera.

KEYWORDS:

Cadmium; Environmental monitoring; Heavy metals; Microbial biosensors; Multiplex; Synthetic biology

PMID:
28319906
DOI:
10.1016/j.bios.2017.03.029
[Indexed for MEDLINE]

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