Design and optimization of a cell-free atrazine biosensor

Adam D. Silverman, Umut Akova, Khalid K. Alam, Michael C. Jewett*, Julius B. Lucks

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Recent advances in cell-free synthetic biology have spurred the development of in vitro molecular diagnostics that serve as effective alternatives to whole-cell biosensors. However, cell-free sensors for detecting manmade organic water contaminants such as pesticides are sparse, partially because few characterized natural biological sensors can directly detect such pollutants. Here, we present a platform for the cell-free detection of one critical water contaminant, atrazine, by combining a previously characterized cyanuric acid biosensor with a reconstituted atrazine-to-cyanuric acid metabolic pathway composed of several protein-enriched bacterial extracts mixed in a one pot reaction. Our cell-free sensor detects atrazine within an hour of incubation at an activation ratio superior to previously reported whole-cell atrazine sensors. We also show that the response characteristics of the atrazine sensor can be tuned by manipulating the component ratios of the cell-free reaction mixture. Our approach of utilizing multiple metabolic steps, encoded in protein-enriched cell-free extracts, to convert a target of interest into a molecule that can be sensed by a transcription factor is modularly designed, which should enable this work to serve as an effective proof-of-concept for rapid field-deployable detection of complex organic water contaminants.

Original languageEnglish (US)
JournalUnknown Journal
DOIs
StatePublished - Sep 24 2019

Keywords

  • Atrazine
  • Biosensor
  • Cell-free
  • Cyanuric acid
  • Metabolism
  • Synthetic biology
  • Transcription factor
  • TX-TL

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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