Development of a bacterial bioassay for atrazine and cyanuric acid detection

Abstract

The s-triazine herbicides are compounds which can disseminate into soils and water. Due to their toxic effects on living organisms, their concentrations in drinking water are legislated by WHO recommendations. Here we have developed for the first time, to the best of our knowledge, an alternative method for physicochemical quantification using two bioluminescent bacterial biosensors: E. coli SM003 for cyanuric acid detection and E. coli SM004 for both atrazine and cyanuric acid detection. The concentration of cyanuric acid detection for E. coli SM003 ranges from 7.83 μM to 2.89 mM, and for E. coli SM004 ranges from 0.22 to 15 μM. Moreover, atrazine detection by E. coli SM004 ranges from 1.08 to 15 μM. According to WHO recommendations, the cyanuric acid detection range is sensitive enough to discriminate between polluted and drinking water. Nevertheless, the detection of atrazine by E. coli SM004 is only applicable for high concentrations of contaminants.

Design of a transcriptional biosensor for the portable, on-demand detection of cyanuric acid

“…Furthermore, even a strong ribosome binding site with a native Pseudomonas promoter gave weak reporter expression suggesting transcriptional incompatibility as the key bottleneck and a possible reason for poor signal in the previously published CYA sensor 27 . Second, hybrid E. coli-Pseudomonas promoters gave strong reporter expression.…”

“…strain ADP1 in E. coli without optimizing for the non-native host. 27 However, poor compatibility of native Pseudomonas promoter with E. coli transcriptional machinery gave low reporter signal and activation, and general design rules could not be inferred from the implementation. To overcome these shortcomings, we sought to use this system to develop a CYA biosensor in E. coli that would enable us to elucidate design rules for activating LTTRs in non-native hosts.…”

A primer on emerging field-deployable synthetic biology tools for global water quality monitoring

“…Both whole-cell and cell-free biosensors have previously been used to detect metals by utilizing natural or engineered proteins; sensors have been reported for cadmium, lead, mercury, arsenic, copper, zinc, nickel, and cobalt, with sensitivities ranging from low parts per million to parts per billion 25,31,82,83 . Sensors for atrazine, a toxic herbicide, have also been developed by encoding a natural metabolic pathway for atrazine's conversion to cyanuric acid, which can be detected with a known protein sensor 84,85 . Furthermore, new cell-free approaches can detect a range of PPCPs, including multiple families of antibiotics and benzalkonium chloride 25,82,86 .…”

Foundations and Emerging Paradigms for Computing in Living Cells

“…Since the design of the bacterial toggle switch and the bacterial oscillator in 2000 [1,2], researchers in the multi-disciplinary field of synthetic biology have developed innovations in the areas of cellular computing [3,4], bio-sensing [5][6][7][8][9][10][11][12][13][14], biochemicals [15][16][17][18][19][20], therapeutics and diagnostics [21][22][23][24][25], pharmaceuticals manufacturing [26][27][28], and biomaterials [29][30][31][32][33][34][35]. With the advent of high-throughput methods to construct and characterize genetic circuits and the continually decreasing costs of DNA synthesis & sequencing [36], synthetic biology is well-poised to continue contributing to areas ranging from answering unsolved questions of biology to generating novel solutions for today's pressing challenges in healthcare and the environment [37].…”