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Sensors 2015, 15(2), 2354-2368; doi:10.3390/s150202354

A High-Throughput Oxidative Stress Biosensor Based on Escherichia coli roGFP2 Cells Immobilized in a k-Carrageenan Matrix

1
Southeast Asia Disaster Prevention Research Initiative (SEADPRI-UKM), Institute for Environment and Development (LESTARI), National University of Malaysia, 43600 Bangi, Selangor, Malaysia
2
Faculty of Science and Technology, National University of Malaysia, 43600 Bangi, Selangor, Malaysia
3
Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan
*
Author to whom correspondence should be addressed.
Received: 14 December 2014 / Revised: 5 January 2015 / Accepted: 14 January 2015 / Published: 22 January 2015
(This article belongs to the Section Biosensors)
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Abstract

Biosensors fabricated with whole-cell bacteria appear to be suitable for detecting bioavailability and toxicity effects of the chemical(s) of concern, but they are usually reported to have drawbacks like long response times (ranging from hours to days), narrow dynamic range and instability during long term storage. Our aim is to fabricate a sensitive whole-cell oxidative stress biosensor which has improved properties that address the mentioned weaknesses. In this paper, we report a novel high-throughput whole-cell biosensor fabricated by immobilizing roGFP2 expressing Escherichia coli cells in a k-carrageenan matrix, for the detection of oxidative stress challenged by metalloid compounds. The E. coli roGFP2 oxidative stress biosensor shows high sensitivity towards arsenite and selenite, with wide linear range and low detection limit (arsenite: 1.0 × 10−3–1.0 × 101 mg·L−1, LOD: 2.0 × 10−4 mg·L−1; selenite: 1.0 × 10−5–1.0 × 102 mg·L−1, LOD: 5.8 × 10−6 mg·L−1), short response times (0–9 min), high stability and reproducibility. This research is expected to provide a new direction in performing high-throughput environmental toxicity screening with living bacterial cells which is capable of measuring the bioavailability and toxicity of environmental stressors in a friction of a second. View Full-Text
Keywords: oxidative stress biosensor; roGFP2; immobilized whole-cell biosensor; redox-sensitive GFP; arsenite; selenite; metalloids; ROS oxidative stress biosensor; roGFP2; immobilized whole-cell biosensor; redox-sensitive GFP; arsenite; selenite; metalloids; ROS
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Ooi, L.; Heng, L.Y.; Mori, I.C. A High-Throughput Oxidative Stress Biosensor Based on Escherichia coli roGFP2 Cells Immobilized in a k-Carrageenan Matrix. Sensors 2015, 15, 2354-2368.

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