Biosensor for branched-chain amino acid metabolism in yeast and applications in isobutanol and isopentanol production. Author Yanfei Zhang, Jeremy Cortez, Sarah Hammer, César Carrasco-López, Sergio Echauri, Jessica Wiggins, Wei Wang, José Avalos Publication Year 2022 Type Journal Article 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. Keywords Leucine, High-Throughput Screening Assays, Biosensing Techniques, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Biosynthetic Pathways, Butanols, Pentanols, Amino Acids, Branched-Chain, Metabolic Engineering, Synthetic Biology, Ethanol, 2-Isopropylmalate Synthase Journal Nat Commun Volume 13 Issue 1 Pages 270 Date Published 2022 Jan 12 ISSN Number 2041-1723 DOI 10.1038/s41467-021-27852-x Alternate Journal Nat Commun PMCID PMC8755756 PMID 35022416 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML