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A Microbial Cocaine Bioreporter.

Anne-Kathrin Grimm1, Dor Rozanes2, Etai Shpigel2

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This summary is machine-generated.

Scientists engineered a novel whole-cell bacterial bioreporter for rapid cocaine detection. This sensitive biosensor offers a promising tool for identifying illegal psychoactive substances in various settings.

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Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Emerging illegal psychoactive compounds challenge existing drug detection methods.
  • Developing specific assays for each new illicit substance is time-consuming and resource-intensive.
  • Whole-cell bacterial bioreporters offer a versatile platform for detecting diverse hazardous chemicals and chemical families.

Purpose of the Study:

  • To develop a microbial bioreporter for the sensitive and specific detection of cocaine, a widely used illegal stimulant.
  • To engineer *Escherichia coli* with a cocaine-responsive bioluminescence system for rapid identification.
  • To optimize the bioreporter's performance and assess its applicability in real-world sample matrices.

Main Methods:

  • Transformation of *Escherichia coli* with a plasmid encoding the *luxCDABEG* bioluminescence genes under the control of a cocaine-responsive promoter.
  • Optimization of the sensing element (*PBen* gene promoter) using accelerated site-directed evolution.
  • Validation of the bioreporter's sensitivity and specificity in buffer solutions and urine samples.
  • Demonstration of cocaine detection using immobilized bioreporter cells on paper strips.

Main Results:

  • The engineered bioreporter demonstrated sensitive cocaine detection with thresholds of 17 ± 8 μg/L in buffer and 130 ± 50 μg/L in urine.
  • Site-directed evolution of the *PBen* promoter enhanced the sensor's performance.
  • The bioreporter system proved effective for cocaine detection in aqueous solutions using immobilized cells on paper strips.

Conclusions:

  • A novel microbial bioreporter system for cocaine detection has been successfully developed.
  • The engineered biosensor exhibits high sensitivity and specificity, offering a valuable tool for drug monitoring.
  • The application of paper strip-based detection highlights the potential for rapid, on-site illicit substance identification.