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A Transcription Factor-Based Biosensor for Detection of Itaconic Acid
Erik K R Hanko1, Nigel P Minton1, Naglis Malys1
1BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, Centre for Biomolecular Sciences , The University of Nottingham , Nottingham , NG7 2RD , United Kingdom.
ACS Synthetic Biology
|April 12, 2018
Summary
Researchers developed a novel itaconate-inducible system for biosensing. This biosensor enables high-throughput screening of microbial strains, facilitating improved itaconic acid (a key platform chemical) biosynthesis.
Area of Science:
- Biotechnology
- Synthetic Biology
- Metabolic Engineering
Background:
- Itaconic acid is a valuable bio-based platform chemical with potential applications in polymer synthesis, aiming to replace petrochemical-derived monomers.
- Current microbial biosynthesis of itaconic acid is hindered by a lack of high-throughput screening methods for strain and condition optimization.
- Identifying novel inducible systems is crucial for advancing microbial production of itaconic acid.
Purpose of the Study:
- To identify and characterize novel itaconate-inducible promoters and transcriptional regulators for microbial applications.
- To develop a robust biosensor for high-throughput screening of itaconic acid production.
- To optimize the expression of key enzymes in the itaconic acid biosynthesis pathway.
Main Methods:
- Identification of LysR-type transcriptional regulators and their cognate promoters from *Yersinia pseudotuberculosis* and *Pseudomonas aeruginosa*.
- Construction and characterization of a fluorescence-based biosensor system (YpItcR/Pccl) in *Escherichia coli* and *Cupriavidus necator*.
- Application of the biosensor to optimize the expression of the *cadA* gene for enhanced itaconate production.
Main Results:
- The YpItcR/Pccl system demonstrated strong inducibility by itaconic acid in *E. coli* (215-fold) and *C. necator* (105-fold).
- The biosensor exhibited dose-dependent detection of itaconate, mesaconate, and cis-/trans-aconitate.
- Fluorescence output correlated well with itaconate concentrations measured by HPLC-UV, validating the biosensor's efficacy.
Conclusions:
- The YpItcR/Pccl inducible system serves as an effective biosensor for itaconic acid.
- This biosensor facilitates high-throughput screening for microbial strain development and optimization of itaconate biosynthesis.
- The developed system holds significant potential for advancing the industrial production of itaconic acid.
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