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Detection of Short-Chain Chlorinated Aliphatic Hydrocarbons through an Engineered Biosensor with Tailored Ligand
Dongdong Chen1, Jiadi Zhao1, Shengmin Xu1
1Information Materials and Intelligent Sensing Laboratory of Anhui Province, Institutes of Physical Science and Information Technology, Anhui University, Hefei 230601, China.
Analytical Chemistry
|September 18, 2024
Summary
A novel whole-cell biosensor was developed for detecting short-chain chlorinated aliphatic hydrocarbons (SCAHs). This biosensor offers enhanced specificity and sensitivity for environmental monitoring of these industrial contaminants.
Area of Science:
- Environmental Science
- Biotechnology
- Analytical Chemistry
Background:
- Short-chain chlorinated aliphatic hydrocarbons (SCAHs) are prevalent industrial chemicals that contaminate aquatic environments, posing risks to ecosystems and human health.
- Whole-cell biosensors offer rapid, cost-effective, real-time monitoring but often suffer from broad ligand specificity.
- Transcriptional factors (TFs) like AlkS are crucial for biosensor development but require optimization for specific analyte detection.
Purpose of the Study:
- To develop a highly specific whole-cell biosensor for the detection of SCAHs.
- To enhance biosensor performance through promoter engineering and directed evolution.
- To validate the biosensor's efficacy in real-world environmental samples.
Main Methods:
- Utilized a semirational transition ligand approach combined with fluorescence-activated cell sorting (FACS) to engineer the AlkS transcriptional factor.
- Employed promoter-directed evolution, modifying constitutive PalkS and inducible PalkB promoters to reduce leakage and improve sensitivity.
- Optimized biosensor M2-463 was characterized for sensitivity, dynamic range, and detection limit.
Main Results:
- Developed an AlkS-based biosensor with high specificity for SCAHs, overcoming limitations of broad TF ligand specificity.
- Achieved an 89% reduction in background fluorescence leakage and a 150-fold increase in fluorescence output after 1 hour of induction.
- The optimized biosensor demonstrated a low detection limit (0.03 ppm) and high recovery rates (95.87–101.20%) in actual water samples.
Conclusions:
- The engineered biosensor M2-463 exhibits superior sensitivity, specificity, and a broad dynamic range for SCAH detection.
- The developed biosensor provides a reliable and accurate tool for environmental monitoring of SCAHs, validated against GC-MS.
- This study presents a promising advancement in whole-cell biosensor technology for practical environmental analysis.
