Journal article
Dietary Change Enables Robust Growth-Coupling of Heterologous Methyltransferase Activity in Yeast
Section for Synthetic Biology, Department of Biotechnology and Biomedicine, Technical University of Denmark1
Computational Biology, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
Department of Biotechnology and Biomedicine, Technical University of Denmark4
University of Washington5
Synthetic Biology Tools for Yeast, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
Department of Environmental Engineering, Technical University of Denmark7
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark8
iLoop, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark9
Genetic modifications of living organisms and proteins are made possible by a catalogue of molecular and synthetic biology tools, yet proper screening assays for genetic variants of interest continue to lag behind. Synthetic growth-coupling (GC) of enzyme activities offers a simple, inexpensive way to track such improvements.
In this follow-up study we present the optimization of a recently established GC design for screening of heterologous methyltransferases (MTases) and related pathways in the yeast Saccharomyces cerevisiae. Specifically, upon testing different media compositions and genetic backgrounds, improved GC of different heterologous MTase activities is obtained.
Furthermore, we demonstrate the strength of the system by screening a library of catechol O-MTase variants converting protocatechuic acid into vanillic acid. We demonstrated high correlation (R2 = 0.775) between vanillic acid and cell density as a proxy for MTase activity. We envision that the improved MTase GC can aid evolution-guided optimization of biobased production processes for methylated compounds with yeast in the future.
Language: | English |
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Publisher: | American Chemical Society |
Year: | 2020 |
Pages: | 3408-3415 |
ISSN: | 21615063 |
Types: | Journal article |
DOI: | 10.1021/acssynbio.0c00348 |
ORCIDs: | Jensen, Michael K. , Hansen, Anne Sofie Lærke and Herrgard, Markus J. |