Decompartmentalization of the yeast mitochondrial metabolism to improve chemical production in Issatchenkia orientalis

Tran, Vinh G.; Tan, Shih-I; Xu, Hao; Weilandt, Daniel R.; Li, Xi; Bhagwat, Sarang S.; Zhu, Zhixin; Guest, Jeremy S.; Rabinowitz, Joshua D.; Zhao, Huimin

doi:10.1038/s41467-025-62304-w

Title: Decompartmentalization of the yeast mitochondrial metabolism to improve chemical production in Issatchenkia orientalis

Journal Article · Sat Aug 02 00:00:00 UTC 2025 · Nature Communications

DOI: https://doi.org/10.1038/s41467-025-62304-w · OSTI ID:2574784

Tran, Vinh G. ^[1]; Tan, Shih-I ^[1];

^[1];

^[2]; Li, Xi ^[2];

^[1]; Zhu, Zhixin ^[1];

^[1];

^[2];

^[1]

University of Illinois Urbana-Champaign, IL (United States)
Princeton University, NJ (United States)

Microbial production of chemicals may suffer from inadequate cofactor provision, a challenge further exacerbated in yeasts due to compartmentalized cofactor metabolism. Here, we perform cofactor engineering through the decompartmentalization of mitochondrial metabolism to improve succinic acid (SA) production in Issatchenkia orientalis. We localize the reducing equivalents of mitochondrial NADH to the cytosol through cytosolic expression of its pyruvate dehydrogenase (PDH) complex and couple a reductive tricarboxylic acid pathway with a glyoxylate shunt, partially bypassing an NADH-dependent malate dehydrogenase to conserve NADH. Cytosolic SA production reaches a titer of 104 g/L and a yield of 0.85 g/g glucose, surpassing the yield of 0.66 g/g glucose constrained by cytosolic NADH availability. Additionally, expressing cytosolic PDH, we expand our I. orientalis platform to enhance acetyl-CoA-derived citramalic acid and triacetic acid lactone production by 1.22- and 4.35-fold, respectively. Our work establishes I. orientalis as a versatile platform to produce markedly reduced and acetyl-CoA-derived chemicals.

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Research Organization:: University of Illinois Urbana-Champaign, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0018420

OSTI ID:: 2574784

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 16; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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