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Screening non-conventional yeasts for acid tolerance and engineering Pichia occidentalis for production of muconic acid

Author

Listed:
  • Michael E. Pyne

    (Concordia University
    Concordia University
    University of Western Ontario)

  • James A. Bagley

    (Concordia University
    Concordia University)

  • Lauren Narcross

    (Concordia University
    Concordia University
    Amyris, Inc.)

  • Kaspar Kevvai

    (Concordia University
    Concordia University
    Pivot Bio)

  • Kealan Exley

    (Concordia University
    Concordia University
    Novo Nordisk Foundation Center for Biosustainability)

  • Meghan Davies

    (Concordia University
    Concordia University
    BenchSci)

  • Qingzhao Wang

    (bp Biosciences Center, San Diego)

  • Malcolm Whiteway

    (Concordia University
    Concordia University)

  • Vincent J. J. Martin

    (Concordia University
    Concordia University)

Abstract

Saccharomyces cerevisiae is a workhorse of industrial biotechnology owing to the organism’s prominence in alcohol fermentation and the suite of sophisticated genetic tools available to manipulate its metabolism. However, S. cerevisiae is not suited to overproduce many bulk bioproducts, as toxicity constrains production at high titers. Here, we employ a high-throughput assay to screen 108 publicly accessible yeast strains for tolerance to 20 g L−1 adipic acid (AA), a nylon precursor. We identify 15 tolerant yeasts and select Pichia occidentalis for production of cis,cis-muconic acid (CCM), the precursor to AA. By developing a genome editing toolkit for P. occidentalis, we demonstrate fed-batch production of CCM with a maximum titer (38.8 g L−1), yield (0.134 g g−1 glucose) and productivity (0.511 g L−1 h−1) that surpasses all metrics achieved using S. cerevisiae. This work brings us closer to the industrial bioproduction of AA and underscores the importance of host selection in bioprocessing.

Suggested Citation

  • Michael E. Pyne & James A. Bagley & Lauren Narcross & Kaspar Kevvai & Kealan Exley & Meghan Davies & Qingzhao Wang & Malcolm Whiteway & Vincent J. J. Martin, 2023. "Screening non-conventional yeasts for acid tolerance and engineering Pichia occidentalis for production of muconic acid," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-41064-5
    DOI: 10.1038/s41467-023-41064-5
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    References listed on IDEAS

    as
    1. Chen Ling & George L. Peabody & Davinia Salvachúa & Young-Mo Kim & Colin M. Kneucker & Christopher H. Calvey & Michela A. Monninger & Nathalie Munoz Munoz & Brenton C. Poirier & Kelsey J. Ramirez & Pe, 2022. "Muconic acid production from glucose and xylose in Pseudomonas putida via evolution and metabolic engineering," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    2. Michael E. Pyne & Kaspar Kevvai & Parbir S. Grewal & Lauren Narcross & Brian Choi & Leanne Bourgeois & John E. Dueber & Vincent J. J. Martin, 2020. "A yeast platform for high-level synthesis of tetrahydroisoquinoline alkaloids," Nature Communications, Nature, vol. 11(1), pages 1-10, December.
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