Cited by 251Microbial Metabolic Engineering and BioproductionBiofuel production and bioconversionFungal and yeast genetics research

Evolutionary Engineering of Saccharomyces cerevisiae for Anaerobic Growth on Xylose

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Sonderegger, Marco, Sauer, Uwe·2003·Applied and Environmental Microbiology

Abstract

Xylose utilization is of commercial interest for efficient conversion of abundant plant material to ethanol. Perhaps the most important ethanol-producing organism, Saccharomyces cerevisiae , however, is incapable of xylose utilization. While S. cerevisiae strains have been metabolically engineered to utilize xylose, none of the recombinant strains or any other naturally occurring yeast has been able to grow anaerobically on xylose. Starting with the recombinant S. cerevisiae strain TMB3001 that overexpresses the xylose utilization pathway from Pichia stipitis , in this study we developed a selection procedure for the evolution of strains that are capable of anaerobic growth on xylose alone. Selection was successful only when organisms were first selected for efficient aerobic growth on xylose alone and then slowly adapted to microaerobic conditions and finally anaerobic conditions, which indicated that multiple mutations were necessary. After a total of 460 generations or 266 days of selection, the culture reproduced stably under anaerobic conditions on xylose and consisted primarily of two subpopulations with distinct phenotypes. Clones in the larger subpopulation grew anaerobically on xylose and utilized both xylose and glucose simultaneously in batch culture, but they exhibited impaired growth on glucose. Surprisingly, clones in the smaller subpopulation were incapable of anaerobic growth on xylose. However, as a consequence of their improved xylose catabolism, these clones produced up to 19% more ethanol than the parental TMB3001 strain produced under process-like conditions from a mixture of glucose and xylose.

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Crossref · 10.1128/aem.69.4.1990-1998.2003

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Authors
Sonderegger, M., & Sauer, U.
Affiliations
ETH Zurich (CH)
Corresponding author
Sauer, Uwe
Venue
Applied and Environmental Microbiology
Published
2003-04
Volume
69
Issue
4
Pages
1990-1998
ISSN
  • 0099-2240 (ISSN-L)
  • 1098-5336
OpenAlex
W2115470699
Cited by
251
References
38
Citations by year
2026: 2 · 2025: 6 · 2024: 2 · 2023: 5 · 2022: 3 · 2021: 4 · 2020: 14 · 2019: 6 · 2018: 8 · 2017: 17 · 2016: 13 · 2015: 5 · 2014: 12 · 2013: 14 · 2012: 18
FWCI
3.68
Type
journal article
Language
en
Open access
Locations
  • Applied and Environmental Microbiology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • European Commission
  • Office Fédéral de l'Education et de la Science
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
  • Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Keywords
Saccharomyces cerevisiae, xylose utilization, anaerobic growth, evolutionary engineering, ethanol production, glucose utilization, xylose catabolism, recombinant yeast strains
MeSH terms
Aldehyde Reductase, Anaerobiosis, Culture Media, Genetic Engineering, Pichia, Recombination, Genetic, Saccharomyces cerevisiae, Sugar Alcohol Dehydrogenases, Xylose, Phosphotransferases (Alcohol Group Acceptor), Evolution, Molecular, D-Xylulose Reductase

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