Project properties

Title Real-time monitoring of metabolites in E. coli using TF-based biosensors
Group Systems and Synthetic Biology
Project type thesis
Credits 36
Supervisor(s) Zihan Gao, Dr. Pieter Candry
Examiner(s) Dr. Pieter Candry
Contact info robert1.smith@wur.nl
Begin date 2025/09/01
End date
Description Real-time monitoring of intracellular metabolites is crucial for understanding and optimizing metabolic processes in E. coli. Transcription factor (TF)-based biosensors, which utilize specific TFs to detect metabolite concentrations, offer a powerful tool for monitoring [1]. They have been widely applied in the detection of target natural products and dynamic regulation of biosynthetic pathways [2].
The goal of this research is to design and validate TF-based biosensors for detecting key metabolites in E. coli. These biosensors will be coupled to fluorescent or other reporters, enabling real-time tracking of metabolic changes. Additionally, the Design-Build-Test-Learn (DBTL) cycle offers a systematic framework that can significantly enhance the performance of biosensors, which has been widely used in manufacturing, metabolic engineering, and entrepreneurship to increase the efficacy and generalizability of biosensors [3]. The MS student will use DBTL cycle(s) to optimize the sensor’s performance towards real application. The results of this project will translate the intracellular concentration of key metabolites into a readable signal and forge a connection between the Design and Learn phases.

References

1. Rogers, J. K., Guzman, C. D., Taylor, N. D., Raman, S., Anderson, K., & Church, G. M. (2015). Synthetic biosensors for precise gene control and real-time monitoring of metabolites. Nucleic acids research, 43(15), 7648–7660. https://doi.org/10.1093/nar/gkv616.

2. Lu, M., Sha, Y., Kumar, V., Xu, Z., Zhai, R., & Jin, M. (2024). Transcription factor-based biosensor: A molecular-guided approach for advanced biofuel synthesis. Biotechnology advances, 72, 108339. https://doi.org/10.1016/j.biotechadv.2024.108339.

3. Chaisupa, P., & Wright, R. C. (2024). State-of-the-art in engineering small molecule biosensors and their applications in metabolic engineering. SLAS technology, 29(2), 100113. https://doi.org/10.1016/j.slast.2023.10.005



Used skills 1) Mastering molecular cloning techniques, including DNA extraction, PCR, plasmid construction and so on.
2) Practical experience with biosensor development and synthetic biology workflows.
Requirements 1) Prefer completed basic molecular microbiology & physiology course. 2) Basic lab skills.