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Here, we report a series of fungal luciferin analogs designed by modification of the arylidene fragment. Electron\u2010donating substituents generally increase activity, whereas extension of\n                    <jats:italic>\u03c0<\/jats:italic>\n                    \u2010conjugation red\u2010shifts emission, with partial decoupling of these effects. These trends parallel those observed for firefly luciferins, indicating that empirical design principles can be transferred between the two systems. Evaluation across fungal luciferases and engineered variant v4, demonstrated broad substrate compatibility and highlighted the combined potential of luciferase engineering and luciferin modification for improving fungal bioluminescence in mammalian bioimaging.\n                  <\/jats:p>","DOI":"10.1002\/cbic.70554","type":"journal-article","created":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T07:15:42Z","timestamp":1791875742000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chemical Tuning of Fungal Bioluminescence in Mammalian Systems"],"prefix":"10.1002","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0417-5812","authenticated-orcid":false,"given":"Anastasia A.","family":"Fadeeva","sequence":"first","affiliation":[{"name":"Department of Biomolecular Chemistry Shemyakin\u2013Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences  Moscow 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