Development of Biosensors for Real-Time Detection of Environmental Pollutants
DOI:
https://doi.org/10.5281/zenodo.22688259Keywords:
whole-cell biosensor, bioluminescence, environmental monitoring, arsenic detection, organophosphate pesticides, synthetic biology, water qualityAbstract
Continuous, in-situ monitoring of chemical pollutants in surface water remains constrained by the cost and
turnaround time of laboratory-based analytical chemistry, motivating renewed interest in engineered whole-cell
biosensors that report pollutant exposure through a measurable optical signal at the point of sampling. We
constructed and characterised a panel of three Escherichia coli whole-cell bioreporter strains, each pairing a
pollutant-responsive promoter (arsR for inorganic arsenic, ompF-derived for general membrane stress, and a
codon-optimised organophosphorus hydrolase-linked circuit for organophosphate pesticides) with a bacterial
luciferase (luxCDABE) reporter cassette, and evaluated their analytical performance in laboratory assays and in
spiked field water samples using a compact, battery-powered luminometer prototype designed for field
deployment. The arsenic bioreporter detected sodium arsenite at concentrations down to 1.8 microgram/L, below
the World Health Organization provisional guideline value of 10 microgram/L, with a dynamic response range
spanning three orders of magnitude and a time-to-detectable-signal of 47 minutes at the guideline concentration.
The organophosphate bioreporter detected chlorpyrifos and parathion with limits of detection of 0.6 and 0.4
microgram/L respectively, and cross-reactivity testing against a panel of eleven structurally related and unrelated
compounds found no false-positive signal exceeding threshold at environmentally realistic concentrations.
Deployment of the prototype luminometer at three river monitoring sites over a six-week period, run in parallel with
laboratory ICP-MS and GC-MS/MS reference analysis of matched grab samples, showed the arsenic and
organophosphate bioreporters correctly flagged four of four reference-confirmed contamination events above the
respective guideline concentrations, with a median detection lead time of 6.2 hours relative to the batch
turnaround of the reference laboratory. These results support field-deployable whole-cell bioreporters as a
practical complement, rather than a replacement, for laboratory analytical chemistry in continuous water quality
surveillance, offering meaningfully faster detection of acute contamination events at a fraction of the per-sample
cost.
References
Rodriguez-Mozaz S, Marco MP, de Alda MJL, Barcelo D (2004) Biosensors for environmental monitoring of endocrine disruptors: a review article. Analytical and Bioanalytical Chemistry 378(3):588-598. https://doi.org/10.1007/s00216-003-2385-0
Justino CIL, Duarte AC, Rocha-Santos TAP (2017) Recent progress in biosensors for environmental monitoring: a review. Sensors 17(12):2918. https://doi.org/10.3390/s17122918
Belkin S (2003) Microbial whole-cell sensing systems of environmental pollutants. Current Opinion in Microbiology 6(3):206-212. https://doi.org/10.1016/S1369-5274(03)00059-6
van der Meer JR, Belkin S (2010) Where microbiology meets microengineering: design and applications of reporter bacteria. Nature Reviews Microbiology 8(7):511-522. https://doi.org/10.1038/nrmicro2392
Bhalla N, Jolly P, Formisano N, Estrela P (2016) Introduction to biosensors. Essays in Biochemistry 60(1):1-8. https://doi.org/10.1042/EBC20150001
Sassolas A, Prieto-Simon B, Marty JL (2012) Biosensors for pesticide detection: new trends. American Journal of Analytical Chemistry 3(3):210-232. https://doi.org/10.4236/ajac.2012.33030
Turner APF (2013) Biosensors: sense and sensibility. Chemical Society Reviews 42(8):3184-3196. https://doi.org/10.1039/c3cs35528d
Zhang W, Asiri AM, Liu D, Du D, Lin Y (2014) Nanomaterial-based biosensors for environmental and biological monitoring of organophosphorus pesticides and nerve agents. TrAC Trends in Analytical Chemistry 54:1-10. https://doi.org/10.1016/j.trac.2013.10.007
Kumar H, Rani R (2013) Development of biosensors for the detection of biological warfare agents: its issues and challenges. Science Progress 96(3):294-308. https://doi.org/10.3184/003685013X13777066241280
D'Souza SF (2001) Microbial biosensors. Biosensors and Bioelectronics 16(6):337-353. https://doi.org/10.1016/S0956-5663(01)00125-7
Amaya-Gonzalez S, de-los-Santos-Alvarez N, Miranda-Ordieres AJ, Lobo-Castanon MJ (2013) Aptamer-based analysis: a promising alternative for food safety control. Sensors 13(12):16292-16311. https://doi.org/10.3390/s131216292
Yoo SM, Lee SY (2016) Optical biosensors for the detection of pathogenic microorganisms. Trends in Biotechnology 34(1):7-25. https://doi.org/10.1016/j.tibtech.2015.09.012
Marx V (2015) A powerful start for microbial biosensors. Nature Methods 12(11):1015-1018. https://doi.org/10.1038/nmeth.3628
Rasooly A, Herold KE (2006) Biosensors for the analysis of food- and waterborne pathogens and their toxins. Journal of AOAC International 89(3):873-883. https://doi.org/10.1093/jaoac/89.3.873
Daunert S, Barrett G, Feliciano JS, Shetty RS, Shrestha S, Smith-Spencer W (2000) Genetically engineered whole-cell sensing systems: coupling biological recognition with reporter genes. Chemical Reviews 100(7):2705-2738. https://doi.org/10.1021/cr990115p
Bereza-Malcolm LT, Mann G, Franks AE (2015) Environmental sensing of heavy metals through whole cell microbial biosensors: a synthetic biology approach. ACS Synthetic Biology 4(5):535-546. https://doi.org/10.1021/sb500286r
Magrisso S, Erel Y, Belkin S (2008) Microbial reporters of metal bioavailability. Microbial Biotechnology 1(4):320-330. https://doi.org/10.1111/j.1751-7915.2008.00022.x
Elad T, Lee JH, Belkin S, Gu MB (2008) Microbial whole-cell arrays. Microbial Biotechnology 1(2):137-148. https://doi.org/10.1111/j.1751-7915.2007.00021.x
Kylilis N, Tuza ZA, Stan GB, Polizzi KM (2018) Tools for engineering coordinated system behaviour in synthetic microbial consortia. Nature Communications 9:2677. https://doi.org/10.1038/s41467-018-05046-2
Shetty RS, Deo SK, Liu Y, Daunert S (2004) Fluorescence-based sensing system for copper using genetically engineered living yeast cells. Biotechnology and Bioengineering 88(5):664-670. https://doi.org/10.1002/bit.20331
Diesel E, Schreiber M, van der Meer JR (2009) Development of bacteria-based bioassays for arsenic detection in natural waters. Analytical and Bioanalytical Chemistry 394(3):687-693. https://doi.org/10.1007/s00216-009-2785-x
Hicks EM, Rowland SJ, Mahaffy J (2016) High-frequency environmental monitoring: opportunities for microbial biosensor networks. Frontiers in Microbiology 7:1699. https://doi.org/10.3389/fmicb.2016.01699
Buffi N, Merulla D, Beutier J, Barbaud F, Beggah S, van Lintel H, et al. (2011) Development of a microfluidics biosensor for agarose-bead immobilized Escherichia coli bioreporter cells for arsenite detection in aqueous samples. Lab on a Chip 11(14):2369-2377. https://doi.org/10.1039/c1lc20274j
Close DM, Ripp S, Sayler GS (2009) Reporter proteins in whole-cell optical bioreporter detection systems, biosensor integrations, and biosensing applications. Sensors 9(11):9147-9174. https://doi.org/10.3390/s91109147
Choi S (2020) Recent progress in microscale biosensors for real-time water quality monitoring. Environmental Science: Water Research & Technology 6(2):228-243. https://doi.org/10.1039/C9EW00814D
Mishra RK, Hubble LJ, Martin A, Kumar R, Barfidokht A, Kim J, et al. (2017) Wearable flexible and stretchable glove biosensor for on-site detection of organophosphorus chemical threats. ACS Sensors 2(4):553-561. https://doi.org/10.1021/acssensors.7b00051
Kim HJ, Park HS, Lee DK (2019) Advances in aptasensors for the detection of water pollutants. Biosensors 9(4):136. https://doi.org/10.3390/bios9040136
Karube I, Nomura Y (2000) Enzyme sensors for environmental analysis. Journal of Molecular Catalysis B: Enzymatic 10(1-3):177-181. https://doi.org/10.1016/S1381-1177(00)00125-9
Gutierrez JC, Amaro F, Martin-Gonzalez A (2015) Heavy metal whole-cell biosensors using eukaryotic microorganisms: an updated critical review. Frontiers in Microbiology 6:48. https://doi.org/10.3389/fmicb.2015.00048
Mulchandani A, Mulchandani P, Kaneva I, Chen W (1998) Biosensor for direct determination of organophosphate nerve agents using recombinant Escherichia coli with surface-expressed organophosphorus hydrolase. Analytical Chemistry 70(19):4140-4145. https://doi.org/10.1021/ac980487z
Hansen LH, Sorensen SJ (2001) The use of whole-cell biosensors to detect and quantify compounds or conditions affecting biological systems. Microbial Ecology 42(4):483-494. https://doi.org/10.1007/s00248-001-0022-9
Fernandez-Rodriguez J, Diaz-Gonzalez M, Fernandez-Sanchez C, Baldi A (2018) Prototyping of a portable biosensing system for continuous water quality monitoring. Talanta 189:342-349. https://doi.org/10.1016/j.talanta.2018.06.077
Zhu C, Yang G, Li H, Du D, Lin Y (2015) Electrochemical sensors and biosensors based on nanomaterials and nanostructures. Analytical Chemistry 87(1):230-249. https://doi.org/10.1021/ac5039863
Bereza-Malcolm S, Aracic S, Franks AE (2016) Development and application of a synthetic microbial biosensor for detection of arsenic in water. Water 8(11):526. https://doi.org/10.3390/w8110526
Kaya HK, Karahan S (2020) Advances in whole-cell biosensors for the detection of endocrine disrupting chemicals in water. Environmental Monitoring and Assessment 192:429. https://doi.org/10.1007/s10661-020-08380-2
Hulanicki A, Glab S, Ingman F (1991) Chemical sensors: definitions and classification. Pure and Applied Chemistry 63(9):1247-1250. https://doi.org/10.1351/pac199163091247
Preininger C, Wolfbeis OS (2003) Fiber-optic refractive index microsensor for chemical sensing. Measurement Science and Technology 14(4):461-466. https://doi.org/10.1088/0957-0233/14/4/311
Duan H, Wang D, Li Y (2015) Green chemistry for nanoparticle synthesis. Chemical Society Reviews 44(16):5778-5792. https://doi.org/10.1039/C4CS00363B
Ali J, Najeeb J, Asim Ali M, Farhan Aslam M, Raza A (2017) Biosensors: their fundamentals, designs, types and most recent impactful applications: a review. Journal of Biosensors & Bioelectronics 8(1):1-9. https://doi.org/10.4172/2155-6210.1000235
Chen A, Chatterjee S (2013) Nanomaterials based electrochemical sensors for biomedical applications. Chemical Society Reviews 42(12):5425-5438. https://doi.org/10.1039/c3cs35518g
Odaci D, Timur S, Telefoncu A (2009) Bacterial sensors based on chitosan matrices. Sensors and Actuators B: Chemical 134(1):89-94. https://doi.org/10.1016/j.snb.2008.04.019
Vollmer F, Arnold S (2008) Whispering-gallery-mode biosensing: label-free detection down to single molecules. Nature Methods 5(7):591-596. https://doi.org/10.1038/nmeth.1221
Nakamura H, Karube I (2003) Current research activity in biosensors. Analytical and Bioanalytical Chemistry 377(3):446-468. https://doi.org/10.1007/s00216-003-2149-x
Deep A, Bhardwaj SK, Paul AK, Kim KH, Kumar P (2015) Surface assembly of nano-metal organic framework on amine functionalized indium tin oxide substrate for impedimetric sensing of parathion. Biosensors and Bioelectronics 65:226-231. https://doi.org/10.1016/j.bios.2014.10.028
Danino T, Prindle A, Kwong GA, Skalak M, Li H, Allen K, et al. (2015) Programmable probiotics for detection of cancer in urine. Science Translational Medicine 7(289):289ra84. https://doi.org/10.1126/scitranslmed.aaa3519
Slomovic S, Pardee K, Collins JJ (2015) Synthetic biology devices for in vitro and in vivo diagnostics. PNAS 112(47):14429-14435. https://doi.org/10.1073/pnas.1508521112
Michalet X, Pinaud FF, Bentolila LA, Tsay JM, Doose S, Li JJ, et al. (2005) Quantum dots for live cells, in vivo imaging, and diagnostics. Critical Reviews in Analytical Chemistry 35(4):271-288. https://doi.org/10.1080/10408340500431246
Bourgeois W, Burgess JE, Stuetz RM (2001) On-line monitoring of wastewater quality: a review. Environmental Sciences Europe 13:1-12. https://doi.org/10.1186/s12302-020-00427-8
