@article {Snoek229922, author = {Tim Snoek and David Romero-Suarez and Jie Zhang and Mette L. Skjoedt and Suresh Sudarsan and Michael K. Jensen and Jay D. Keasling}, title = {An orthogonal and pH-tunable sensor-selector for muconic acid biosynthesis in yeast}, elocation-id = {229922}, year = {2017}, doi = {10.1101/229922}, publisher = {Cold Spring Harbor Laboratory}, 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 the sensor-selector does not affect production, and find that tuning pH of the cultivation medium limits the rise of non-producing 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 produced 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.}, URL = {https://www.biorxiv.org/content/early/2017/12/06/229922}, eprint = {https://www.biorxiv.org/content/early/2017/12/06/229922.full.pdf}, journal = {bioRxiv} }