Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast

Mette Louise Skjødt, Tim Snoek, Kanchana Rueksomtawin Kildegaard, Dushica Arsovska, M. Eichenberger, T. J. Goedecke, Arun Stephen Rajkumar, Jie Zhang, Mette Kristensen, B. Lehka, Solvej Siedler, Irina Borodina, Michael Krogh Jensen, Jay Keasling

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Whole-cell biocatalysts have proven a tractable path toward sustainable production of bulk and fine chemicals. Yet the screening of libraries of cellular designs to identify best-performing biocatalysts is most often a low-throughput endeavor. For this reason, the development of biosensors enabling real-time monitoring of production has attracted attention. Here we applied systematic engineering of multiple parameters to search for a general biosensor design in the budding yeast Saccharomyces cerevisiae based on small-molecule binding transcriptional activators from the prokaryote superfamily of LysR-type transcriptional regulators (LTTRs). We identified a design supporting LTTR-dependent activation of reporter gene expression in the presence of cognate small-molecule inducers. As proof of principle, we applied the biosensors for in vivo screening of cells producing naringenin or cis,cis-muconic acid at different levels, and found that reporter gene output correlated with production. The transplantation of prokaryotic transcriptional activators into the eukaryotic chassis illustrates the potential of a hitherto untapped biosensor resource useful for biotechnological applications.
Original languageEnglish
JournalNature Chemical Biology
Volume12
Issue number11
Pages (from-to)951-958
Number of pages10
ISSN1552-4450
DOIs
Publication statusPublished - 2016

Keywords

  • Metabolic engineering
  • Model invertebrates
  • Synthetic biology
  • Transcription

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