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

TitleBiosensor for branched-chain amino acid metabolism in yeast and applications in isobutanol and isopentanol production.
Publication TypeJournal Article
Year of Publication2022
AuthorsZhang, Y, Cortez, JD, Hammer, SK, Carrasco-López, C, Echauri, SÁGarcía, Wiggins, JB, Wang, W, Avalos, JL
JournalNat Commun
Volume13
Issue1
Pagination270
Date Published2022 01 12
ISSN2041-1723
Keywords2-Isopropylmalate Synthase, Amino Acids, Branched-Chain, Biosensing Techniques, Biosynthetic Pathways, Butanols, Ethanol, High-Throughput Screening Assays, Leucine, Metabolic Engineering, Pentanols, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Synthetic Biology
Abstract

<p>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.</p>

DOI10.1038/s41467-021-27852-x
Alternate JournalNat Commun
PubMed ID35022416
PubMed Central IDPMC8755756
Grant ListT32 GM007388 / GM / NIGMS NIH HHS / United States