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A High Throughput Screen for Biomining Cellulase Activity from Metagenomic Libraries
Published on: February 1, 2011
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Engineering a biosensor based high-throughput screening platform for high-yield caffeic acid production in
Daoguang Tian1, Zhen Qin2, Weilin Liu1
1School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, China; Key Laboratory of Systems Bioengineering, Ministry of Education (Tianjin University), Tianjin, 300072, China; Zhejiang Institute of Tianjin University, Shaoxing, 312300, China.
Metabolic Engineering
|October 1, 2025
Summary
We developed a biosensor-driven platform to efficiently produce caffeic acid (CA) in E. coli. This method overcomes production challenges, achieving the highest reported CA titer to date.
Area of Science:
- Microbial biotechnology
- Metabolic engineering
- Synthetic biology
Background:
- Caffeic acid (CA) is a valuable phenolic compound with diverse applications.
- Microbial production of CA is hindered by low enzyme activity and product toxicity.
Purpose of the Study:
- To develop an efficient high-throughput screening (HTS) platform for caffeic acid (CA) production.
- To overcome bottlenecks in CA biosynthesis, including enzyme activity and product cytotoxicity.
Main Methods:
- Engineered a novel p-coumaric acid (p-CA) biosensor using the CarR transcription factor.
- Integrated the biosensor with fluorescence-activated cell sorting (FACS) for HTS.
- Performed metabolic engineering on E. coli strains for enhanced CA production.
Main Results:
- Optimized p-CA biosensor with improved sensitivity and dynamic range.
- Identified an improved FjTALG85S mutant with 6.85-fold higher catalytic activity.
- Achieved a record CA titer of 9.61 g/L in a 5-L bioreactor using engineered strain CA8.
Conclusions:
- The developed biosensor-HTS platform accelerates the engineering of microbial cell factories.
- This work provides a powerful tool for producing p-CA and other derived chemicals efficiently.
- Key challenges in CA biosynthesis, such as enzyme efficiency and cytotoxicity, were successfully addressed.
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