Erik K.R. Hanko
A transcription factor-based biosensor for detection of itaconic acid
Hanko, Erik K.R.; Minton, Nigel P.; Malys, Naglis
Authors
Professor NIGEL MINTON nigel.minton@nottingham.ac.uk
Professor of Applied Molecular Microbiology
Naglis Malys
Abstract
Itaconic acid is an important platform chemical that can easily be incorporated into polymers and has the potential to replace petrochemical-based acrylic or methacrylic acid. A number of microorganisms have been developed for the biosynthesis of itaconate including Aspergillus terreus, Escherichia coli and Saccharomyces cerevisiae. However, the number of strains and conditions that can be tested for increased itaconate titers are currently limited due to the lack of high-throughput screening methods. Here we identified itaconate-inducible promoters and their corresponding LysR-type transcriptional regulators from Yersinia pseudotuberculosis and Pseudomonas aeruginosa. We show that the YpItcR/Pccl inducible system is highly inducible by itaconic acid in the model gammaproteobacterium E. coli and the betaproteobacterium Cupriavidus necator (215- and 105-fold, respectively). The kinetics and dynamics of the YpItcR/Pccl inducible system are investigated and we demonstrate, that in addition to itaconate, the genetically encoded biosensor is capable of detecting mesaconate, cis-, and trans-aconitate in a dose-dependent manner. Moreover, the fluorescence-based biosensor is applied in E. coli to identify the optimum expression level of cadA, the product of which catalyzes the conversion of cis-aconitate into itaconate. The fluorescence output is shown to correlate well with itaconate concentrations quantified using high-performance liquid chromatography coupled with ultraviolet spectroscopy. This work highlights the potential of the YpItcR/Pccl inducible system to be applied as biosensor for high-throughput microbial strain development to facilitate improved itaconate biosynthesis.
Citation
Hanko, E. K., Minton, N. P., & Malys, N. (2018). A transcription factor-based biosensor for detection of itaconic acid. ACS Synthetic Biology, 7(5), 1436–1446. https://doi.org/10.1021/acssynbio.8b00057
Journal Article Type | Article |
---|---|
Acceptance Date | Apr 10, 2018 |
Publication Date | Apr 11, 2018 |
Deposit Date | Apr 16, 2018 |
Publicly Available Date | Aug 16, 2018 |
Journal | ACS Synthetic Biology |
Electronic ISSN | 2161-5063 |
Publisher | American Chemical Society |
Peer Reviewed | Peer Reviewed |
Volume | 7 |
Issue | 5 |
Pages | 1436–1446 |
DOI | https://doi.org/10.1021/acssynbio.8b00057 |
Public URL | https://nottingham-repository.worktribe.com/output/924365 |
Publisher URL | https://pubs.acs.org/doi/10.1021/acssynbio.8b00057 |
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Publisher Licence URL
https://creativecommons.org/licenses/by/4.0/
Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf
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