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

Yanfei Zhang, Jeremy D. Cortez, Sarah K. Hammer, César Carrasco-López, Sergio Á. García Echauri, Jessica B. Wiggins, Wei Wang and José L. Avalos ()
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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

Nature Communications, 2022, vol. 13, issue 1, 1-14

Abstract: 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.

Date: 2022
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DOI: 10.1038/s41467-021-27852-x

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