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Epigenetic modulation by suppressing epigenetic inhibitors, such as 5-azacytidine (5-aza), has been shown to activate silent biosynthetic gene clusters within a fungus and causes the production of novel secondary metabolites. This research examined this epigenetic modification strategy in the poorly studied filamentous fungus, <jats:italic>Ceratorhiza hydrophila<\/jats:italic>, which may help induce the additional production of bioactive compounds.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>The results from genomic and spectroscopic analyses (ISSR profiling and FTIR spectroscopy) indicated that 50 \u00b5M 5-aza produced substantial global DNA demethylation and genomic changes in <jats:italic>C. hydrophila<\/jats:italic> with no impact on cell viability. The epigenetic changes associated with the DNA demethylation prompted a notable and selective change in antimicrobial profile to suppress antibacterial activity against strains such as <jats:italic>Clostridium sporogenes<\/jats:italic> while also showing a robust induction of antifungal activity against <jats:italic>Candida albicans<\/jats:italic> (22\u00a0mm inhibition zone). GC-MS was performed for a deep-dive characterization of the metabolic profile which revealed, for example, a dramatic alteration of the profile including production of new secondary metabolites such as a novel indole derivative and diisooctyl phthalate, which did not exist in the untreated control. In silico analyses, such as modelling the promoter and molecular docking opportunities, offered a believable mechanistic rationale for the effects seen, linked to the predicted modulation of primary biosynthetic pathways.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>This study demonstrates that epigenetic modulation can be used to successfully unlock latent biosynthetic capability in <jats:italic>C. hydrophila<\/jats:italic> resulting in the production of unique compounds with strong and selective antifungal activity. These results demonstrate the advantages of epigenetic screening of unique fungal sources in the search for new drug leads.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/s12866-025-04330-8","type":"journal-article","created":{"date-parts":[[2025,9,9]],"date-time":"2025-09-09T02:34:50Z","timestamp":1757385290000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Epigenetic modulation of Ceratorhiza hydrophila by 5-azacytidine enhances antifungal metabolite production: insights from antimicrobial, metabolic, genomic and computational analyses"],"prefix":"10.1186","volume":"25","author":[{"given":"Rehab M.","family":"Abdelhamid","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3728-7431","authenticated-orcid":false,"given":"Elham R. 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