Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast

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

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

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.
OriginalsprogEngelsk
TidsskriftNature Chemical Biology
Vol/bind12
Udgave nummer11
Sider (fra-til)951-958
ISSN1552-4450
DOI
StatusUdgivet - 2016

Emneord

  • SACCHAROMYCES-CEREVISIAE
  • SYNTHETIC BIOLOGY
  • MAMMALIAN-CELLS
  • GENE-EXPRESSION
  • SINORHIZOBIUM-MELILOTI
  • BIOSYNTHETIC PATHWAYS
  • MUCONIC ACID
  • CYC1 GENE
  • PROTEIN
  • OPTIMIZATION

Citer dette

Skjødt, M. L., Snoek, T., Kildegaard, K. R., Arsovska, D., Eichenberger, M., Goedecke, T. J., ... Keasling, J. (2016). Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast. Nature Chemical Biology, 12(11), 951-958. https://doi.org/10.1038/NCHEMBIO.2177
Skjødt, Mette Louise ; Snoek, Tim ; Kildegaard, Kanchana Rueksomtawin ; Arsovska, Dushica ; Eichenberger, Michael ; Goedecke, Tobias J. ; Rajkumar, Arun Stephen ; Zhang, Jie ; Kristensen, Mette ; Lehka, Beata Joanna ; Siedler, Solvej ; Borodina, Irina ; Jensen, Michael Krogh ; Keasling, Jay. / Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast. I: Nature Chemical Biology. 2016 ; Bind 12, Nr. 11. s. 951-958.
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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.",
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author = "Skj{\o}dt, {Mette Louise} and Tim Snoek and Kildegaard, {Kanchana Rueksomtawin} and Dushica Arsovska and Michael Eichenberger and Goedecke, {Tobias J.} and Rajkumar, {Arun Stephen} and Jie Zhang and Mette Kristensen and Lehka, {Beata Joanna} and Solvej Siedler and Irina Borodina and Jensen, {Michael Krogh} and Jay Keasling",
year = "2016",
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pages = "951--958",
journal = "Nature Chemical Biology",
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Skjødt, ML, Snoek, T, Kildegaard, KR, Arsovska, D, Eichenberger, M, Goedecke, TJ, Rajkumar, AS, Zhang, J, Kristensen, M, Lehka, BJ, Siedler, S, Borodina, I, Jensen, MK & Keasling, J 2016, 'Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast', Nature Chemical Biology, bind 12, nr. 11, s. 951-958. https://doi.org/10.1038/NCHEMBIO.2177

Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast. / Skjødt, Mette Louise; Snoek, Tim; Kildegaard, Kanchana Rueksomtawin; Arsovska, Dushica; Eichenberger, Michael ; Goedecke, Tobias J. ; Rajkumar, Arun Stephen; Zhang, Jie; Kristensen, Mette; Lehka, Beata Joanna; Siedler, Solvej; Borodina, Irina; Jensen, Michael Krogh; Keasling, Jay.

I: Nature Chemical Biology, Bind 12, Nr. 11, 2016, s. 951-958.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

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, Michael

AU - Goedecke, Tobias J.

AU - Rajkumar, Arun Stephen

AU - Zhang, Jie

AU - Kristensen, Mette

AU - Lehka, Beata Joanna

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 - SACCHAROMYCES-CEREVISIAE

KW - SYNTHETIC BIOLOGY

KW - MAMMALIAN-CELLS

KW - GENE-EXPRESSION

KW - SINORHIZOBIUM-MELILOTI

KW - BIOSYNTHETIC PATHWAYS

KW - MUCONIC ACID

KW - CYC1 GENE

KW - PROTEIN

KW - OPTIMIZATION

KW - SACCHAROMYCES-CEREVISIAE

KW - SYNTHETIC BIOLOGY

KW - MAMMALIAN-CELLS

KW - GENE-EXPRESSION

KW - SINORHIZOBIUM-MELILOTI

KW - BIOSYNTHETIC PATHWAYS

KW - MUCONIC ACID

KW - CYC1 GENE

KW - PROTEIN

KW - OPTIMIZATION

U2 - 10.1038/NCHEMBIO.2177

DO - 10.1038/NCHEMBIO.2177

M3 - Journal article

VL - 12

SP - 951

EP - 958

JO - Nature Chemical Biology

JF - Nature Chemical Biology

SN - 1552-4450

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ER -

Skjødt ML, Snoek T, Kildegaard KR, Arsovska D, Eichenberger M, Goedecke TJ et al. Engineering prokaryotic transcriptional activators as metabolite biosensors in yeast. Nature Chemical Biology. 2016;12(11):951-958. https://doi.org/10.1038/NCHEMBIO.2177