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

Michael E. Pyne, James A. Bagley, Lauren Narcross, Kaspar Kevvai, Kealan Exley, Meghan Davies, Qingzhao Wang, Malcolm Whiteway and Vincent J. J. Martin ()
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Michael E. Pyne: Concordia University
James A. Bagley: Concordia University
Lauren Narcross: Concordia University
Kaspar Kevvai: Concordia University
Kealan Exley: Concordia University
Meghan Davies: Concordia University
Qingzhao Wang: bp Biosciences Center, San Diego
Malcolm Whiteway: Concordia University
Vincent J. J. Martin: Concordia University

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-41064-5

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