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

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

doi:10.1038/s41467-021-27852-x

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

Journal Article · 11 January 2022 · Nature Communications

DOI: https://doi.org/10.1038/s41467-021-27852-x · OSTI ID:1839492

Zhang, Yanfei; Cortez, Jeremy D.; Hammer, Sarah K.;

; García Echauri, Sergio Á.; Wiggins, Jessica B.;

;

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.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: NONE; SC0019363

OSTI ID:: 1839492

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 13; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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