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Multicolored Living Biosensors for Highly Selective and Ultrasensitive Detection of Aromatic Pollutants in Water
CC BY 4.0 · Indian J Med Paediatr Oncol 2024; 45(S 01): S1-S16
DOI: DOI: 10.1055/s-0044-1788221
*Corresponding author contact: (e-mail: arindam@chem.iitb.ac.in/ruchi@chem.iitb.ac.in).
Abstract
Background: Carcinogenic mono-aromatic phenols and the benzene, toluene, ethylbenzene, and m-xylene pollutants pose health risks even at low ppb levels in drinking water. Detecting them for water quality surveillance is challenging. This study fine-tuned the phenol sensor, MopR, through structure-guided mutagenesis, creating whole-cell biosensors (WCBs) leveraging synthetic biology for multi-compound sensing.
Material and Method: Utilizing cloning strategies, an array of fluorescent sensor cells was generated. Performance was evaluated via fluorescence spectroscopy at both bulk and single bacterial cell levels, utilizing a wide-field microscope. To enhance shelf-life and portability, the cells were lyophilized and coupled with cost-effective, simple imaging method, for on-site pollutant sensing.
Results: The first-generation sensor cells encountered challenges such as background fluorescence and insufficient detection sensitivity. To address these, synthetic transcriptional and translational modules were integrated, enabling detection of specific compounds down to drinking water limits (< ~1 ppb). Alterations to the reporter gene GFP module engineered a series of visible-range sensors, allowing simultaneous and specific detection of four pollutants in water samples. Integrating quantitative estimation of aromatic pollutants from images, with a portable, field-deployable, modified foldscope setup, resulted in a rapid, user-friendly, and cost-effective on-site monitoring tool for detecting aromatic pollutants in drinking water.
Conclusion: The amalgamation of structural biology principles and synthetically enabled engineered cell design has led to the development of novel, cost-effective and portable array of biosensors for effective detection of phenols and benzene and its derivatives sensitively in drinking water sources, which is a major global threat.
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
*Corresponding author contact: (e-mail: arindam@chem.iitb.ac.in/ruchi@chem.iitb.ac.in).
Abstract
Background: Carcinogenic mono-aromatic phenols and the benzene, toluene, ethylbenzene, and m-xylene pollutants pose health risks even at low ppb levels in drinking water. Detecting them for water quality surveillance is challenging. This study fine-tuned the phenol sensor, MopR, through structure-guided mutagenesis, creating whole-cell biosensors (WCBs) leveraging synthetic biology for multi-compound sensing.
Material and Method: Utilizing cloning strategies, an array of fluorescent sensor cells was generated. Performance was evaluated via fluorescence spectroscopy at both bulk and single bacterial cell levels, utilizing a wide-field microscope. To enhance shelf-life and portability, the cells were lyophilized and coupled with cost-effective, simple imaging method, for on-site pollutant sensing.
Results: The first-generation sensor cells encountered challenges such as background fluorescence and insufficient detection sensitivity. To address these, synthetic transcriptional and translational modules were integrated, enabling detection of specific compounds down to drinking water limits (< ~1 ppb). Alterations to the reporter gene GFP module engineered a series of visible-range sensors, allowing simultaneous and specific detection of four pollutants in water samples. Integrating quantitative estimation of aromatic pollutants from images, with a portable, field-deployable, modified foldscope setup, resulted in a rapid, user-friendly, and cost-effective on-site monitoring tool for detecting aromatic pollutants in drinking water.
Conclusion: The amalgamation of structural biology principles and synthetically enabled engineered cell design has led to the development of novel, cost-effective and portable array of biosensors for effective detection of phenols and benzene and its derivatives sensitively in drinking water sources, which is a major global threat.
No conflict of interest has been declared by the author(s).
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India