Dietary Change Enables Robust Growth-Coupling of Heterologous Methyltransferase Activity in Yeast

Anne Sofie Lærke Hansen, Maitreya J. Dunham, Dushica Arsovska, Jie Zhang, Jay D. Keasling, Markus J. Herrgard, Michael K. Jensen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

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.
Original languageEnglish
JournalACS Synthetic Biology
Volume9
Pages (from-to)3408-3415
Number of pages8
ISSN2161-5063
DOIs
Publication statusPublished - 2020

Keywords

  • Growth-coupling
  • Methyltransferase
  • Yeast
  • Synthetic biology
  • Natural products
  • Screening

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