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A High Throughput Screen for Biomining Cellulase Activity from Metagenomic Libraries
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Engineering Modular and Highly Sensitive Cell-Based Biosensors for Aromatic Contaminant Monitoring and

Shengwei Sun1, Kailin Peng1, Sen Sun1

  • 1Key Laboratory of Food Processing and Quality Control, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China.

ACS Synthetic Biology
|February 23, 2023
PubMed
Summary

Engineered bacterial biosensors with novel genetic circuits significantly enhance signal amplification for detecting environmental contaminants. This breakthrough improves sensitivity for identifying phenolic compounds and screening valuable enzymes from soil samples.

Keywords:
DmpR-based biosensoraromatic contaminant detectiongenetic circuitintracellular enzyme screeningsignal amplifying

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Area of Science:

  • Synthetic Biology
  • Environmental Biotechnology
  • Microbial Engineering

Background:

  • Existing whole-cell biosensors often lack the sensitivity required for practical applications.
  • Signal amplification is crucial for improving the performance of biosensor systems.
  • Phenolic compounds are common environmental contaminants requiring robust detection methods.

Purpose of the Study:

  • To develop novel modular genetic circuits for signal amplification in whole-cell biosensors.
  • To enhance the sensitivity and detection capabilities for phenolic contaminants.
  • To apply the improved biosensor for screening functional enzymes in environmental samples.

Main Methods:

  • Construction of serial and parallel modular genetic circuits in *Escherichia coli*.
  • Engineering biosensors using the σ54-dependent phenol-responsive regulator DmpR and GFP reporter.
  • Testing biosensor sensitivity, detection limits, and ranges for phenolic compounds and enzyme screening.

Main Results:

  • Serial and parallel circuits achieved 9- and 16-fold higher sensitivity, respectively.
  • Rapid detection of six phenolic contaminants within 12 hours.
  • Low limits of detection for benzopyrene (2.5 ppb) and tetracycline (2.2 ppb).
  • Successful screening of soil metagenomic libraries, identifying novel esterase enzymes with 73% positive rate.

Conclusions:

  • The developed genetic circuits provide a powerful signal amplification strategy for cell-based biosensors.
  • This approach significantly enhances environmental contaminant detection and enzyme discovery.
  • The biosensor system offers a versatile platform for environmental monitoring and biocatalyst screening.