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Biosensor for branched-chain amino acid metabolism in yeast and applications in isobutanol and isopentanol production

Author

Listed:
  • Yanfei Zhang

    (Princeton University)

  • Jeremy D. Cortez

    (Princeton University)

  • Sarah K. Hammer

    (Princeton University)

  • César Carrasco-López

    (Princeton University)

  • Sergio Á. García Echauri

    (Princeton University)

  • Jessica B. Wiggins

    (Princeton University)

  • Wei Wang

    (Princeton University)

  • José L. Avalos

    (Princeton University
    Princeton University
    Princeton University
    Princeton University)

Abstract

Branched-chain amino acid (BCAA) metabolism fulfills numerous physiological roles and can be harnessed to produce valuable chemicals. However, the lack of eukaryotic biosensors specific for BCAA-derived products has limited the ability to develop high-throughput screens for strain engineering and metabolic studies. Here, we harness the transcriptional regulator Leu3p from Saccharomyces cerevisiae to develop a genetically encoded biosensor for BCAA metabolism. In one configuration, we use the biosensor to monitor yeast production of isobutanol, an alcohol derived from valine degradation. Small modifications allow us to redeploy Leu3p in another biosensor configuration that monitors production of the leucine-derived alcohol, isopentanol. These biosensor configurations are effective at isolating high-producing strains and identifying enzymes with enhanced activity from screens for branched-chain higher alcohol (BCHA) biosynthesis in mitochondria as well as cytosol. Furthermore, this biosensor has the potential to assist in metabolic studies involving BCAA pathways, and offers a blueprint to develop biosensors for other products derived from BCAA metabolism.

Suggested Citation

  • Yanfei Zhang & Jeremy D. Cortez & Sarah K. Hammer & César Carrasco-López & Sergio Á. García Echauri & Jessica B. Wiggins & Wei Wang & José L. Avalos, 2022. "Biosensor for branched-chain amino acid metabolism in yeast and applications in isobutanol and isopentanol production," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-021-27852-x
    DOI: 10.1038/s41467-021-27852-x
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    Cited by:

    1. Charlotte Cautereels & Jolien Smets & Peter Bircham & Dries De Ruysscher & Anna Zimmermann & Peter De Rijk & Jan Steensels & Anton Gorkovskiy & Joleen Masschelein & Kevin J. Verstrepen, 2024. "Combinatorial optimization of gene expression through recombinase-mediated promoter and terminator shuffling in yeast," Nature Communications, Nature, vol. 15(1), pages 1-17, December.

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