Oxygenation influences xylose fermentation and gene expression in the yeast genera Spathaspora and Scheffersomyces

Barros, Katharina O.; Mader, Megan; Krause, David J.; Pangilinan, Jasmyn; Andreopoulos, Bill; Lipzen, Anna; Mondo, Stephen J.; Grigoriev, Igor V.; Rosa, Carlos A.; Sato, Trey K.; Hittinger, Chris Todd

doi:10.1186/s13068-024-02467-8

Title: Oxygenation influences xylose fermentation and gene expression in the yeast genera Spathaspora and Scheffersomyces

Journal Article · Wed Feb 07 00:00:00 EST 2024 · Biotechnology for Biofuels and Bioproducts

DOI:https://doi.org/10.1186/s13068-024-02467-8· OSTI ID:2325391

; Mader, Megan; Krause, David J.;

;

Abstract Background Cost-effective production of biofuels from lignocellulose requires the fermentation of d -xylose. Many yeast species within and closely related to the genera Spathaspora and Scheffersomyces (both of the order Serinales) natively assimilate and ferment xylose. Other species consume xylose inefficiently, leading to extracellular accumulation of xylitol. Xylitol excretion is thought to be due to the different cofactor requirements of the first two steps of xylose metabolism. Xylose reductase (XR) generally uses NADPH to reduce xylose to xylitol, while xylitol dehydrogenase (XDH) generally uses NAD ⁺ to oxidize xylitol to xylulose, creating an imbalanced redox pathway. This imbalance is thought to be particularly consequential in hypoxic or anoxic environments. Results We screened the growth of xylose-fermenting yeast species in high and moderate aeration and identified both ethanol producers and xylitol producers. Selected species were further characterized for their XR and XDH cofactor preferences by enzyme assays and gene expression patterns by RNA-Seq. Our data revealed that xylose metabolism is more redox balanced in some species, but it is strongly affected by oxygen levels. Under high aeration, most species switched from ethanol production to xylitol accumulation, despite the availability of ample oxygen to accept electrons from NADH. This switch was followed by decreases in enzyme activity and the expression of genes related to xylose metabolism, suggesting that bottlenecks in xylose fermentation are not always due to cofactor preferences. Finally, we expressed XYL genes from multiple Scheffersomyces species in a strain of Saccharomyces cerevisiae . Recombinant S. cerevisiae expressing XYL1 from Scheffersomyces xylosifermentans , which encodes an XR without a cofactor preference, showed improved anaerobic growth on xylose as the primary carbon source compared to S. cerevisiae strain expressing XYL genes from Scheffersomyces stipitis . Conclusion Collectively, our data do not support the hypothesis that xylitol accumulation occurs primarily due to differences in cofactor preferences between xylose reductase and xylitol dehydrogenase; instead, gene expression plays a major role in response to oxygen levels. We have also identified the yeast Sc. xylosifermentans as a potential source for genes that can be engineered into S. cerevisiae to improve xylose fermentation and biofuel production.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF); USDA NIFA

Grant/Contract Number:: AC02-05CH11231; SC0018409; DEB-2110403

OSTI ID:: 2325391

Alternate ID(s):: OSTI ID: 2335323

Journal Information:: Biotechnology for Biofuels and Bioproducts, Journal Name: Biotechnology for Biofuels and Bioproducts Vol. 17 Journal Issue: 1; ISSN 2731-3654

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Title: Oxygenation influences xylose fermentation and gene expression in the yeast genera Spathaspora and Scheffersomyces

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