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This review integrates the latest research progress on the regulation of fungal ergosterol biosynthesis and its role in drug resistance and pathogenicity. We comprehensively discuss the functions of key enzymes (such as Erg11p\/Cyp51A, Erg6p, Erg3p, and Erg25p) in the mevalonate, late, and alternative pathways. Notably, we highlight the complex regulation of cyp51A expression by factors such as SrbA, AtrR, CBC, HapX, and NCT in Aspergillus fumigatus, and elucidate the distinctive roles of Upc2, Adr1, and Rpn4 in Candida species. Importantly, we summarize recent discoveries on the CprA-dependent regulation of Cyp51A\/Erg11p and heme-mediated stability control. Based on these findings, we propose innovative antifungal strategies, including dual-target inhibition and multi-enzyme inhibition by natural products, which provide novel insights and potential directions for the development of next-generation antifungal therapies.<\/jats:p>","DOI":"10.3390\/microorganisms13040862","type":"journal-article","created":{"date-parts":[[2025,4,10]],"date-time":"2025-04-10T11:26:41Z","timestamp":1744284401000},"page":"862","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":49,"title":["Regulation of Ergosterol Biosynthesis in Pathogenic Fungi: Opportunities for Therapeutic Development"],"prefix":"10.3390","volume":"13","author":[{"given":"Lingyun","family":"Song","sequence":"first","affiliation":[{"name":"The Key Laboratory of Biotechnology for Medicinal and Edible Plant Resources of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sha","family":"Wang","sequence":"additional","affiliation":[{"name":"Huzhou Key Laboratory of Precise Prevention and Control of Major Chronic Diseases, Huzhou University, Huzhou 313000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hang","family":"Zou","sequence":"additional","affiliation":[{"name":"Antibiotics Research and Re-Evaluation Key Laboratory of Sichuan Province, Chengdu University, Chengdu 610059, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaokang","family":"Yi","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Biotechnology for Medicinal and Edible Plant Resources of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shihan","family":"Jia","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Biotechnology for Medicinal and Edible Plant Resources of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8623-9068","authenticated-orcid":false,"given":"Rongpeng","family":"Li","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Biotechnology for Medicinal and Edible Plant Resources of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1374-1353","authenticated-orcid":false,"given":"Jinxing","family":"Song","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Biotechnology for Medicinal and Edible Plant Resources of Jiangsu Province, School of Life Sciences, Jiangsu Normal University, Xuzhou 221116, China"},{"name":"Qian Xuesen Collaborative Research Center of Astrochemistry and Space Life Science, Institute of Drug Discovery Technology, Ningbo 315211, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Lass-Fl\u00f6rl, C., Dietl, A.M., Kontoyiannis, D.P., and Brock, M. 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