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Engineering Yeast Transcription Machinery for Improved Ethanol Tolerance and Production
Abstract
Global transcription machinery engineering (gTME) is an approach for reprogramming gene transcription to elicit cellular phenotypes important for technological applications. Here we show the application of gTME to Saccharomyces cerevisiae for improved glucose/ethanol tolerance, a key trait for many biofuels programs. Mutagenesis of the transcription factor Spt15p and selection led to dominant mutations that conferred increased tolerance and more efficient glucose conversion to ethanol. The desired phenotype results from the combined effect of three separate mutations in the SPT15 gene [serine substituted for phenylalanine (Phe 177 Ser) and, similarly, Tyr 195 His, and Lys 218 Arg]. Thus, gTME can provide a route to complex phenotypes that are not readily accessible by traditional methods.
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“For example, laboratory selection can lead to large-scale rewiring of gene regulatory networks, but an approach, that the authors called gTME, global transcription machinery engineering, can also yield similar results [2].”
From Adaptive laboratory evolution—Principles and applications for biotechnology (2013) · cites this paper as [2]
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“Global transcription machinery engineering (gTME) has been successfully used to enhance the tolerance of S. cerevisiae to ethanol (Alper et al. 2006 ).”
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Sahoo, R. K., Das, A., Gaur, M., Dey, S., Sahoo, S., & Subudhi, E. (2019) · Journal of Pure and Applied Microbiology · doi:10.22207/jpam.13.3.13 · Cited by 6
Sazegari, S., Niazi, A., Zinati, Z., & Eskandari, M. H. (2022) · PLoS ONE · doi:10.1371/journal.pone.0259476 · Cited by 6
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Exploring the Capabilities of the Geobiosphere's Microbial Genome
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“This approach has already been employed in yeast to improve ethanol tolerance (Alper et al. 2006); however, only a standard laboratory yeast strain was used in this work.”
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Guo, Y. Q., Dong, Y., Hong, X., Pang, X., Chen, D., & Chen, X. (2016) · International Journal of Molecular Sciences · doi:10.3390/ijms17111813 · Cited by 6
“Mutagenesis of the transcription factor Spt15p led to dominant mutations that conferred higher tolerance and more efficient glucose conversion to ethanol [12].”
BackgroundMethodgTME for Construction of Recombinant Yeast Co-fermenting Xylose and Glucose
Liu, H., Xu, L., YAN, M., LAI, C., & Ouyang, P. (2008) · Chinese journal of biotechnology/Shengwu gongcheng xuebao · doi:10.1016/s1872-2075(08)60048-5 · Cited by 6
“发生变化从而改变或提高目标基因的转录及表 达([6])。gTME 方法特别能够操纵大肠杆菌的σ因子和 在酿酒酵母中的 TFIID 组分引起转录子多基因的重 排。TFII D 由 TBP(TATA 结合蛋白)和十几种 TBP 相关因子(TAF)构成([7])。转录因子 spt15 属于转录起 始复合物部分, 是一种 TATA 结合蛋白, 与 RNA 聚”
Rational design for over-production of desirable microbial metabolites by precision engineering
Gao, H. C., Zhou, X., Gou, Z., Zhuo, Y., Fu, C., Liu, M., Song, F., Ashforth, E. J., & Zhang, L. (2010) · Antonie van Leeuwenhoek · doi:10.1007/s10482-010-9442-4 · Cited by 6
“Recently, global perturbations in gene expression has been accomplished by engineering cellular transcription and translation machinery (Alper et al. 2006) by a tool termed global transcription machinery engineering (gTME).”
BackgroundTolerance of Microbial Biocatalysts to Feedstocks, Products, and Environmental Conditions
Huang, M., Peabody, G. L., V, & Kao, K. C. (2016) · Springer eBooks · doi:10.1007/978-3-319-41966-4_5 · Cited by 6
Xu, Y.-Y., Wang, L., Hu, W., Xue, J., Xiao, W., Zhang, H., Zhang, J., & Chen, X. (2025) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2025.09.017 · Cited by 5
A cellular platform for production of C 4 monomers
Davis, M. A., Yu, V. Y., Fu, B., Wen, M., Koleski, E. J., Silverman, J., Berdan, C. A., Nomura, D. K., & Chang, M. C. Y. (2023) · Chemical Science · doi:10.1039/d3sc02773b · Cited by 5
Evolutionary Engineering for Industrial Microbiology
Vanee, N., Fisher, A. B., & Fong, S. S. (2012) · Sub-cellular biochemistry/Subcellular biochemistry · doi:10.1007/978-94-007-5055-5_3 · Cited by 5
Metadata
- Authors
- Alper, H., Moxley, J., Nevoigt, E., Fink, G. R., & Stephanopoulos, G.
- Affiliations
- Whitehead Institute for Biomedical Research (US)
- Corresponding author
- Stephanopoulos, Gregory
- Venue
- Science
- Published
- 2006-12-08
- Volume
- 314
- Issue
- 5805
- Pages
- 1565-1568
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2080537888
- Cited by
- 823
- References
- 26
- Citations by year
- 2026: 10 · 2025: 23 · 2024: 31 · 2023: 18 · 2022: 28 · 2021: 39 · 2020: 43 · 2019: 37 · 2018: 39 · 2017: 44 · 2016: 49 · 2015: 61 · 2014: 55 · 2013: 57 · 2012: 68
- FWCI
- 14.67
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (GM035010)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Keywords
- Saccharomyces cerevisiae, ethanol tolerance, glucose fermentation, transcription factor mutations, dominant mutations, biofuel production, yeast strain engineering
- MeSH terms
- Ethanol, Fermentation, Genetic Engineering, Glucose, Phenotype, Saccharomyces cerevisiae, Transcription Factors, Transcription, Genetic, Transformation, Genetic, Up-Regulation, Gene Expression Regulation, Fungal, Mutagenesis, Cell Cycle Proteins, Amino Acid Substitution, Gene Expression Profiling, Saccharomyces cerevisiae Proteins, TATA-Box Binding Protein, Transcription Factor TFIID, TATA-Binding Protein Associated Factors
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