Abstract
Microbes offer enormous potential for production of industrially relevant chemicals and therapeutics, yet the rapid identification of high-producing microbes from large genetic libraries is a major bottleneck in modern cell factory development. Here, we develop and apply a synthetic selection system in Saccharomyces cerevisiae that couples the concentration of muconic acid, a plastic precursor, to cell fitness by using the prokaryotic transcriptional regulator BenM driving an antibiotic resistance gene. We show that the sensor-selector does not affect production nor fitness, and find that tuning pH of the cultivation medium limits the rise of nonproducing cheaters. We apply the sensor-selector to selectively enrich for best-producing variants out of a large library of muconic acid production strains, and identify an isolate that produces more than 2 g/L muconic acid in a bioreactor. We expect that this sensor-selector can aid the development of other synthetic selection systems based on allosteric transcription factors.
Original language | English |
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Journal | A C S Synthetic Biology |
Volume | 7 |
Issue number | 4 |
Pages (from-to) | 995-1003 |
ISSN | 2161-5063 |
DOIs | |
Publication status | Published - 2018 |
Keywords
- Biosensor
- Evolution
- Metabolic engineering
- Sustainability
- Transcriptional activator
- Yeast