Journal article
An Orthogonal and pH-Tunable Sensor-Selector for Muconic Acid Biosynthesis in Yeast
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Synthetic Biology Tools for Yeast, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
iLoop, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
Pre-Pilot Plant, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
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.
Language: | English |
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Publisher: | American Chemical Society |
Year: | 2018 |
Pages: | 995-1003 |
ISSN: | 21615063 |
Types: | Journal article |
DOI: | 10.1021/acssynbio.7b00439 |
ORCIDs: | Snoek, Tim , Romero-Suarez, David , Zhang, Jie , Ambri, Francesca and Jensen, Michael K. |
Bacterial Proteins BenM protein, Acinetobacter Bioreactors Biosensing Techniques Drug Resistance, Fungal High-Throughput Screening Assays Hydrogen-Ion Concentration Metabolic Engineering Microorganisms, Genetically-Modified Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Sorbic Acid Transcription Factors biosensor evolution metabolic engineering muconic acid sustainability transcriptional activator yeast