1887

Abstract

The global use of organophosphate insecticides (OPPs) and the growing concern of off-target side effects due to OPP exposure has prompted the need for sensitive and economical detection methods. Here we set out to engineer a previously identified OPP responsive transcription factor, ChpR, from to respond to alternative OPPs and generate a repertoire of whole-cell biosensors for OPPs. The ChpR transcription factor and cognate promoter P have been shown to activate transcription in the presence of the OPP chlorpyrifos (CPF). Utilizing a GFP reporter regulated by ChpR in a whole-cell biosensor we found that the system responds significantly better to 3,5,6-trichloro-2-pyridinol (TCP), the main degradation product of CPF, compared to CPF itself. This biosensor was able to respond to TCP at 390 nM within 4 h compared to 50 µM of CPF in 7 h. The ChpR-P, and the activating ligand TCP, were able to regulate expression of a kanamycin resistance/sucrose sensitivity () selection/counterselection module suitable for high throughput mutagenesis screening studies. The ability to control both GFP and the module demonstrates the utility of this reporter for the detection of CPF affected areas. The ChpR-P system serves as an additional positive regulator switch to add to the growing repertoire of controllers available within synthetic biology.

Funding
This study was supported by the:
  • Defense Threat Reduction Agency
    • Principle Award Recipient: HenryS Gibbons
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/acmi/10.1099/acmi.0.000297
2021-12-15
2022-01-27
Loading full text...

Full text loading...

/deliver/fulltext/acmi/3/12/acmi000297.html?itemId=/content/journal/acmi/10.1099/acmi.0.000297&mimeType=html&fmt=ahah

References

  1. Gardner GT, Stern PC. Environmental Problems and Human Behavior Allyn & Bacon; 1996
    [Google Scholar]
  2. Aktar MW, Sengupta D, Chowdhury A. Impact of pesticides use in agriculture: their benefits and hazards. Interdiscip Toxicol 2009; 2:1–12 [View Article] [PubMed]
    [Google Scholar]
  3. Ali I, Jain C. Groundwater contamination and health hazards by some of the most commonly used pesticides. Curr Sci 1998; 75:1011–1014
    [Google Scholar]
  4. Aroonvilairat S, Tangjarukij C, Sornprachum T, Chaisuriya P, Siwadune T et al. Effects of topical exposure to a mixture of chlorpyrifos, cypermethrin and captan on the hematological and immunological systems in male wistar rats. Environ Toxicol Pharmacol 2018; 59:60 [View Article] [PubMed]
    [Google Scholar]
  5. Barron MG, Woodburn KB. Ecotoxicology of chlorpyrifos. Rev Environ Contam Toxicol 1995; 144:1–93 [View Article] [PubMed]
    [Google Scholar]
  6. Beketov MA, Kefford BJ, Schäfer RB, Liess M. Pesticides reduce regional biodiversity of stream invertebrates. Proc Natl Acad Sci U S A 2013; 110:11039–11043 [View Article] [PubMed]
    [Google Scholar]
  7. Chakraborty S, Mukherjee S, Roychoudhury S, Siddique S, Lahiri T et al. Chronic exposures to cholinesterase-inhibiting pesticides adversely affect respiratory health of agricultural workers in India. J Occup Health 2009; 51:488–497 [View Article] [PubMed]
    [Google Scholar]
  8. Clune AL, Ryan PB, Barr DB. Have regulatory efforts to reduce organophosphorus insecticide exposures been effective?. Environ Health Perspect 2012; 120:521–525 [View Article] [PubMed]
    [Google Scholar]
  9. Eddleston M, Buckley NA, Eyer P, Dawson AH. Management of acute organophosphorus pesticide poisoning. The Lancet 2008; 371:597–607 [View Article]
    [Google Scholar]
  10. Morgan MK, Sheldon LS, Croghan CW, Jones PA, Robertson GL et al. Exposures of preschool children to chlorpyrifos and its degradation product 3,5,6-trichloro-2-pyridinol in their everyday environments. J Expo Anal Environ Epidemiol 2005; 15:297–309 [View Article] [PubMed]
    [Google Scholar]
  11. Stachniuk A, Fornal E. Liquid chromatography-mass spectrometry in the analysis of pesticide residues in food. Food Anal Methods 2015; 9:1654–1665 [View Article]
    [Google Scholar]
  12. Verma N, Bhardwaj A. Biosensor technology for pesticides--a review. Appl Biochem Biotechnol 2015; 175:3093–3119 [View Article] [PubMed]
    [Google Scholar]
  13. Gui Q, Lawson T, Shan S, Yan L, Liu Y. The application of whole cell-based biosensors for use in environmental analysis and in medical diagnostics. Sensors 2017; 17:1623 [View Article]
    [Google Scholar]
  14. Karig DK. Cell-free synthetic biology for environmental sensing and remediation. Curr Opin Biotechnol 2017; 45:69–75 [View Article] [PubMed]
    [Google Scholar]
  15. Fernandez-López R, Ruiz R, de la Cruz F, Moncalián G. Transcription factor-based biosensors enlightened by the analyte. Front Microbiol 2015; 6:648 [View Article] [PubMed]
    [Google Scholar]
  16. Whangsuk W, Dubbs JM, Sallabhan R, Somsongkul K, Mongkolsuk S et al. ChpR is a chlorpyrifos-responsive transcription regulator in Sinorhizobium meliloti. J Mol Microbiol Biotechnol 2010; 18:141–147 [View Article] [PubMed]
    [Google Scholar]
  17. Whangsuk W, Thiengmag S, Dubbs J, Mongkolsuk S, Loprasert S. Specific detection of the pesticide chlorpyrifos by a sensitive genetic-based whole cell biosensor. Anal Biochem 2016; 493:11–13 [View Article] [PubMed]
    [Google Scholar]
  18. Chouichit P, Whangsuk W, Sallabhan R, Mongkolsuk S, Loprasert S. A highly sensitive biosensor with a single-copy evolved sensing cassette for chlorpyrifos pesticide detection. Microbiology (Reading) 2020; 166:1019–1024 [View Article] [PubMed]
    [Google Scholar]
  19. Whangsuk W, Dubbs JM, Sallabhan R, Somsongkul K, Mongkolsuk S et al. ChpR is a chlorpyrifos-responsive transcription regulator in Sinorhizobium meliloti. J Mol Microbiol Biotechnol 2010; 18:141–147 [View Article] [PubMed]
    [Google Scholar]
  20. Racke KD. Environmental fate of chlorpyrifos. Rev Environ Contam Toxicol 1993; 131:1–150 [View Article] [PubMed]
    [Google Scholar]
  21. Racke KD, Steele KP, Yoder RN, Dick WA, Avidov E. Factors affecting the hydrolytic degradation of chlorpyrifos in soil. J Agric Food Chem 1996; 44:1582–1592 [View Article]
    [Google Scholar]
  22. Garmendia J, de las Heras A, Galvão TC, de Lorenzo V. Tracing explosives in soil with transcriptional regulators of pseudomonas putida evolved for responding to nitrotoluenes. Microb Biotechnol 2008; 1:236–246 [View Article] [PubMed]
    [Google Scholar]
  23. Tomlin CDS. The Pesticide Manual, A World Compendium, 14th ed. Alton, Hampshire, United Kingdom: British Crop Protection Council; 2006
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/acmi/10.1099/acmi.0.000297
Loading
/content/journal/acmi/10.1099/acmi.0.000297
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error