Engineering Bacillus Spores to Display Nicotine Oxidase: In Situ Specific and Sensitive Nicotine Detection
Xisheng Xiong1,2,3, Shuangsheng Xiong4, Ruiqi Zhu1,2,3
1State Key Laboratory of Natural Medicines, China Pharmaceutical University, 639 Longmian Avenue, Jiangning, Nanjing 211198, China.
ACS Sensors
|May 7, 2025
Summary
This study developed a novel live electrochemical biosensor for highly sensitive nicotine detection. The biosensor integrates engineered enzymes on bacterial spores with advanced nanomaterials for real-time monitoring.
Area of Science:
- Biotechnology
- Materials Science
- Electrochemistry
Background:
- Nicotine exposure presents a significant public health concern.
- Highly sensitive and real-time detection methods for nicotine are crucial.
- Existing methods may lack sensitivity or real-time capabilities.
Purpose of the Study:
- To develop a novel live electrochemical biosensor for sensitive and real-time nicotine detection.
- To integrate synthetic biology and materials engineering for enhanced biosensor performance.
- To validate the proof-of-concept for using engineered enzymes in electrochemical sensing.
Main Methods:
- Genomic integration of a genetically engineered NOX mutant onto Bacillus subtilis spores.
- Fabrication of a hierarchical porous FeSe@MHCS composite electrode.
- Utilizing H2O2-mediated signal amplification for nicotine detection via electrochemical techniques.
Main Results:
- Achieved an ultralow detection limit of 0.25 nM for nicotine.
- Demonstrated minimal matrix interference in blood/urine samples (98-107% recovery).
- Exhibited exceptional stability (98% retention after 14 days) and reproducibility (RSD 0.62%).
Conclusions:
- The developed live electrochemical biosensor is effective for real-time nicotine monitoring.
- The integration of spore-displayed enzymes and nanomaterials offers a robust biosensing platform.
- This work provides a blueprint for developing enzyme-based electrochemical biosensors for diverse analytes.
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