Lipid membrane remodeling and metabolic response during isobutanol and ethanol exposure in Zymomonas mobilis
Open accessAbstract
Background: Recent engineering efforts have targeted the ethanologenic bacterium Zymomonas mobilis for isobutanol production. However, significant hurdles remain due this organism’s vulnerability to isobutanol toxicity, adversely affecting its growth and productivity. The limited understanding of the physiological impacts of isobutanol on Z. mobilis constrains our ability to overcome these production barriers. Results: We utilized a systems-level approach comprising LC–MS/MS-based lipidomics, metabolomics, and shotgun proteomics, to investigate how exposure to ethanol and isobutanol impact the lipid membrane composition and overall physiology of Z. mobilis . Our analysis revealed significant and distinct alterations in membrane phospholipid and fatty acid composition resulting from ethanol and isobutanol exposure. Notably, ethanol exposure increased membrane cyclopropane fatty acid content and expression of cyclopropane fatty acid (CFA) synthase. Surprisingly, isobutanol decreased cyclopropane fatty acid content despite robust upregulation of CFA synthase. Overexpression of the native Z. mobilis’ CFA synthase increased cyclopropane fatty acid content in all phospholipid classes and was associated with a significant improvement in growth rates in the presence of added ethanol and isobutanol. Heterologous expression of CFA synthase from Clostridium acetobutylicum resulted in a near complete replacement of unsaturated fatty acids with cyclopropane fatty acids, affecting all lipid classes. However, this did not translate to improved growth rates under isobutanol exposure. Correlating with its greater susceptibility to isobutanol, Z. mobilis exhibited more pronounced alterations in its proteome, metabolome, and overall cell morphology—including cell swelling and formation of intracellular protein aggregates —when exposed to isobutanol compared to ethanol. Isobutanol triggered a broad stress response marked by the upregulation of heat shock proteins, efflux transporters, DNA repair systems, and the downregulation of cell motility proteins. Isobutanol also elicited widespread dysregulation of Z. mobilis ’ primary metabolism evidenced by increased levels of nucleotide degradation intermediates and the depletion of biosynthetic and glycolytic intermediates. Conclusions: This study provides a comprehensive, systems-level evaluation of the impact of ethanol and isobutanol exposure on the lipid membrane composition and overall physiology of Z. mobilis . These findings will guide engineering of Z. mobilis towards the creation of isobutanol-tolerant strains that can serve as robust platforms for the industrial production of isobutanol from lignocellulosic sugars.
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Ethanol exposure leads to increased cyclopropane fatty acid content and upregulation of CFA synthase in Z. mobilis.
“Our findings revealed distinct responses to these stressors: ethanol exposure leads to increased cyclopropane fatty acid (CFA) content and upregulation of CFA synthase, whereas isobutanol exposure decreases CFA content despite robust CFA synthase expression.”
From Physiological and metabolic responses of <i>Zymomonas mobilis</i> to lignocellulosic hydrolysate (2025) · cites this paper as [13]
Isobutanol exposure decreases cyclopropane fatty acid content in Z. mobilis despite robust CFA synthase expression.
“Our findings revealed distinct responses to these stressors: ethanol exposure leads to increased cyclopropane fatty acid (CFA) content and upregulation of CFA synthase, whereas isobutanol exposure decreases CFA content despite robust CFA synthase expression.”
From Physiological and metabolic responses of <i>Zymomonas mobilis</i> to lignocellulosic hydrolysate (2025) · cites this paper as [13]
Isobutanol causes widespread metabolic dysregulation in Z. mobilis, evident from increased nucleotide degradation intermediates and depletion of biosynthetic and glycolytic intermediates.
“Additionally, isobutanol causes widespread metabolic dysregulation, evident from increased nucleotide degradation intermediates and depletion of biosynthetic and glycolytic intermediates.”
From Physiological and metabolic responses of <i>Zymomonas mobilis</i> to lignocellulosic hydrolysate (2025) · cites this paper as [13]
Isobutanol triggers a broad stress response in Z. mobilis characterized by upregulation of heat shock proteins, efflux transporters, and DNA repair systems, along with downregulation of cell motility proteins.
“Isobutanol also triggers a broad stress response, characterized by the upregulation of heat shock proteins, efflux transporters, and DNA repair systems, along with the downregulation of cell motility proteins.”
From Physiological and metabolic responses of <i>Zymomonas mobilis</i> to lignocellulosic hydrolysate (2025) · cites this paper as [13]
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- Authors
- Rivera Vazquez, J., Trujillo, E., Williams, J., She, F., Getahun, F., Callaghan, M. M., Coon, J. J., & Amador-Noguez, D.
- Affiliations
- University of Wisconsin–Madison (US)
- Venue
- Biotechnology for Biofuels and Bioproducts
- Published
- 2024-01-28
- Volume
- 17
- Issue
- 1
- Pages
- 14-14
- ISSN
- 2731-3654 (ISSN-L)
- PMC
- PMC10823705
- OpenAlex
- W4391291448
- Cited by
- 13
- References
- 84
- Citations by year
- 2026: 8 · 2025: 5
- FWCI
- 1.76
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Biotechnology for Biofuels and Bioproducts (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- OSTI OAI (U.S. Department of Energy Office of Scientific and Technical Information) (submittedVersion)
- Funders
- U.S. Department of Energy (DE-SC0018409)
- National Institutes of Health (P41GM108538)
- 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 › Biochemistry › Lipid metabolism and biosynthesis
- Keywords
- Zymomonas mobilis, ethanol exposure, cyclopropane fatty acids, cyclopropane fatty acid synthase, membrane phospholipids, fatty acid composition, stress response, heat shock proteins, efflux pumps, nucleotide degradation, primary metabolism, cell morphology, protein aggregation, isobutanol production
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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