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Evolutionary engineering of mixed-sugar utilization by a xylose-fermenting strain
Abstract
We have recently reported about a Saccharomyces cerevisiae strain that, in addition to the Piromyces XylA xylose isomerase gene, overexpresses the native genes for the conversion of xylulose to glycolytic intermediates. This engineered strain (RWB 217) exhibited unprecedentedly high specific growth rates and ethanol production rates under anaerobic conditions with xylose as the sole carbon source. However, when RWB 217 was grown on glucose-xylose mixtures, a diauxic growth pattern was observed with a relatively slow consumption of xylose in the second growth phase. After prolonged cultivation in an anaerobic, xylose-limited chemostat, a culture with improved xylose uptake kinetics was obtained. This culture also exhibited improved xylose consumption in glucose-xylose mixtures. A further improvement in mixed-sugar utilization was obtained by prolonged anaerobic cultivation in automated sequencing-batch reactors on glucose-xylose mixtures. A final single-strain isolate (RWB 218) rapidly consumed glucose-xylose mixtures anaerobically, in synthetic medium, with a specific rate of xylose consumption exceeding 0.9 gg(-1)h(-1). When the kinetics of zero trans-influx of glucose and xylose of RWB 218 were compared to that of the initial strain, a twofold higher capacity (V(max)) as well as an improved K(m) for xylose was apparent in the selected strain. It is concluded that the kinetics of xylose fermentation are no longer a bottleneck in the industrial production of bioethanol with yeast.
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“Mutations in acrAB and marC were found independently in these studies; further changes were linked to carbon and nitrogen metabolism via gatY and tnaA[86], enterobactin synthesis [63] or attenuated rpoS activity via hfq mutations [87].”
From Adaptive laboratory evolution—Principles and applications for biotechnology (2013) · cites this paper as [87]
In the citing paper
“Moreover, mutations identified in isobutanol-resistant strains showed a high degree of epistasis [87].”
From Adaptive laboratory evolution—Principles and applications for biotechnology (2013) · cites this paper as [87]
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Metadata
- Authors
- KUYPER, M., TOIRKENS, M., DIDERICH, J., WINKLER, A., VANDIJKEN, J., & PRONK, J.
- Affiliations
- Delft University of Technology (NL)
- Corresponding author
- PRONK, J
- ORCID
- Venue
- FEMS Yeast Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2005-07
- Volume
- 5
- Issue
- 10
- Pages
- 925-934
- ISSN
- 1567-1356 (ISSN-L)
- OpenAlex
- W2116734717
- Cited by
- 341
- References
- 46
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 8 · 2023: 3 · 2022: 6 · 2021: 4 · 2020: 14 · 2019: 8 · 2018: 12 · 2017: 24 · 2016: 23 · 2015: 11 · 2014: 28 · 2013: 29 · 2012: 28
- FWCI
- 7.36
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- FEMS Yeast Research (publishedVersion)
- PubMed (publishedVersion)
- 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
- xylose fermentation, Saccharomyces cerevisiae, xylose isomerase, diauxic growth, ethanol production, bioethanol, anaerobic cultivation, sequencing batch reactor, chemostat
- MeSH terms
- Anaerobiosis, Biotechnology, Carbohydrates, Culture Media, Fermentation, Saccharomyces cerevisiae, Time Factors, Xylose
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