Antifungal Activity of Disalt of Epipyrone A from Epicoccum nigrum Likely via Disrupted Fatty Acid Elongation and Sphingolipid Biosynthesis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Fungal Strains
2.3. Extraction and Purification of Disalt of Epipyrone A (DEA)
2.4. Liquid Bioactivity Assay
2.5. Chemical-Genetic Analysis of Haploid Deletion Library
2.6. Culture Preparation for Metabolomics
2.7. Sampling and Extraction Procedure for Intracellular Metabolite Analysis
2.8. Extracellular Metabolite Analysis
2.9. Separation, Identification, and Analysis of Metabolites
2.10. Proteome-Wide Protein Abundance and Localization Analysis
2.11. Thin-Layer Chromatography (TLC)
3. Results
3.1. Bioativity of DEA in Yeast
3.2. Chemical-Genetic Profiling Distinguishes the Importance of Endocytosis, Fatty Acid Elongation, Cell Wall Integrity, and Actin Cytoskeleton as Mechanisms Buffering DEA Bioactivity
3.3. Metabolomic Analysis Reveals Increased Membrane Permeability and Increased Intracellular Phenolic Acid Accumulation Induced by DEA
3.4. Fluorescence Microscopy Confirms Interruption of Sphingolipid Biosynthesis by DEA
3.5. Thin Layer Chromatography Confirms the Effect of DEA on Lipid Metabolism
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Lee, A.J.; Hammond, J.; Sheridan, J.; Swift, S.; Munkacsi, A.B.; Villas-Boas, S.G. Antifungal Activity of Disalt of Epipyrone A from Epicoccum nigrum Likely via Disrupted Fatty Acid Elongation and Sphingolipid Biosynthesis. J. Fungi 2024, 10, 597. https://doi.org/10.3390/jof10090597
Lee AJ, Hammond J, Sheridan J, Swift S, Munkacsi AB, Villas-Boas SG. Antifungal Activity of Disalt of Epipyrone A from Epicoccum nigrum Likely via Disrupted Fatty Acid Elongation and Sphingolipid Biosynthesis. Journal of Fungi. 2024; 10(9):597. https://doi.org/10.3390/jof10090597
Chicago/Turabian StyleLee, Alex J., Joseph Hammond, Jeffrey Sheridan, Simon Swift, Andrew B. Munkacsi, and Silas G. Villas-Boas. 2024. "Antifungal Activity of Disalt of Epipyrone A from Epicoccum nigrum Likely via Disrupted Fatty Acid Elongation and Sphingolipid Biosynthesis" Journal of Fungi 10, no. 9: 597. https://doi.org/10.3390/jof10090597