TY - JOUR
T1 - Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast
AU - Skjødt, Mette Louise
AU - Snoek, Tim
AU - Kildegaard, Kanchana Rueksomtawin
AU - Arsovska, Dushica
AU - Eichenberger, M.
AU - Goedecke, T. J.
AU - Rajkumar, Arun Stephen
AU - Zhang, Jie
AU - Kristensen, Mette
AU - Lehka, B.
AU - Siedler, Solvej
AU - Borodina, Irina
AU - Jensen, Michael Krogh
AU - Keasling, Jay
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
KW - Metabolic engineering
KW - Model invertebrates
KW - Synthetic biology
KW - Transcription
U2 - 10.1038/nchembio.2177
DO - 10.1038/nchembio.2177
M3 - Journal article
C2 - 27642864
SN - 1552-4450
VL - 12
SP - 951
EP - 958
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 11
ER -