Journal article
Regulatory control circuits for stabilizing long-term anabolic product formation in yeast
Synthetic Biology Tools for Yeast, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Technical University of Denmark3
Delft University of Technology4
Pre-Pilot Plant, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
Bacterial Synthetic Biology, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark7
Engineering living cells for production of chemicals, enzymes and therapeutics can burden cells due to use of limited native co-factor availability and/or expression burdens, totalling a fitness deficit compared to parental cells encoded through long evolutionary trajectories to maximise fitness. Ultimately, this discrepancy puts a selective pressure against fitness-burdened engineered cells under prolonged bioprocesses, and potentially leads to complete eradication of high-performing engineered cells at the population level.
Here we present the mutation landscapes of fitness-burdened yeast cells engineered for vanillin-β-glucoside production. Next, we design synthetic control circuits based on transcriptome analysis and biosensors responsive to vanillin-β-glucoside pathway intermediates in order to stabilize vanillin-β-glucoside production over ~55 generations in sequential passage experiments.
Furthermore, using biosensors with two different modes of action we identify control circuits linking vanillin-β-glucoside pathway flux to various essential cellular functions, and demonstrate control circuits robustness and almost 2-fold higher vanillin-β-glucoside production, including 5-fold increase in total vanillin-β-glucoside pathway metabolite accumulation, in a fed-batch fermentation compared to vanillin-β-glucoside producing cells without control circuits.
Language: | English |
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Year: | 2020 |
Pages: | 369-380 |
ISSN: | 10967176 and 10967184 |
Types: | Journal article |
DOI: | 10.1016/j.ymben.2020.07.006 |
ORCIDs: | 0000-0002-7766-2040 , 0000-0002-6563-2792 , ter Horst, Jolanda , Ambri, Francesca , Rugbjerg, Peter , 0000-0002-5311-1404 , D'ambrosio, Vasil , Sommer, Morten Otto Alexander and Jensen, Michael Krogh |