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 journalArticle

5 Scopus citations

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 ratios of enriched extracts in 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 modular. Our work thus serves as an effective proof-of-concept for a scheme of "metabolic biosensing", which should enable rapid, field-deployable detection of complex organic water contaminants.

Original languageEnglish (US)
Pages (from-to)671-677
Number of pages7
JournalACS synthetic biology
Volume9
Issue number3
DOIs
StatePublished - Mar 20 2020

Keywords

  • TX-TL
  • atrazine
  • biosensor
  • cell-free
  • cyanuric acid
  • metabolism
  • synthetic biology
  • transcription factor

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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