title: Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast.
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.
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Mette L
authors.initials: ML
authors.lastname: Skjoedt
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Tim
authors.initials: T
authors.lastname: Snoek
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Kanchana R
authors.initials: KR
authors.lastname: Kildegaard
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Dushica
authors.initials: D
authors.lastname: Arsovska
authors.affiliation: Evolva SA, Reinach, Switzerland.|Department of Biology, Technical University Darmstadt, Darmstadt, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Eichenberger
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Tobias J
authors.initials: TJ
authors.lastname: Goedecke
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Arun S
authors.initials: AS
authors.lastname: Rajkumar
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Mette
authors.initials: M
authors.lastname: Kristensen
authors.affiliation: Evolva Biotech A/S, Copenhagen, Denmark.|Department of Science and Environment, Roskilde University, Roskilde, Denmark.
authors.forename: Beata J
authors.initials: BJ
authors.lastname: Lehka
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Solvej
authors.identifier: http://orcid.org/0000-0002-5445-8792
authors.initials: S
authors.lastname: Siedler
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Irina
authors.identifier: http://orcid.org/0000-0002-8452-1393
authors.initials: I
authors.lastname: Borodina
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.
authors.forename: Michael K
authors.identifier: http://orcid.org/0000-0001-7574-4707
authors.initials: MK
authors.lastname: Jensen
authors.affiliation: The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark.|Joint BioEnergy Institute, Emeryville, California, USA.|Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.|Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, California, USA.|Department of Bioengineering, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Jay D
authors.initials: JD
authors.lastname: Keasling
mesh_terms: D015374:Biosensing Techniques; D011387:Prokaryotic Cells / Q000378:metabolism*; D015202:Protein Engineering; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D014157:Transcription Factors