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Therefore, this study evaluated efficacy of enzyme cocktail containing lettuce\u2010expressed chitinases, glucanases and mannanase against <jats:italic>C. albicans<\/jats:italic> in cell culture or cancer patient samples. Site\u2010specific integration of the Man1, CelO, Cbh1, Cbh2 expression cassettes and removal of the selectable marker gene from lettuce transplastome was confirmed using three sets of PCR primers. Homoplasmy in transplastomic lines was confirmed in Southern blots by the absence of untransformed genomes. Unlike tobacco, lettuce transplastomic lines had no mosaic phenotype and yielded more biomass than untransformed plants. Maternal inheritance of transgenes was confirmed by lack of segregation when seedlings were germinated in the selection medium. Mannanase expressed in lettuce chloroplasts is thermostable, exhibiting the highest activity at 70\u2009\u00b0C; expression increased up to 70\u2009days of growth and is active in a broad range of pH (5\u20138). Endoglucanases and exoglucanases expressed in lettuce chloroplasts have a broad range of activities between 30 and 75\u2009\u00b0C and pH 4\u20138. Based on observed biomass and mannanase expression level, up to 8640 million enzyme units could be harvested per acre\/year. The enzyme cocktail effectively degraded the fungal cell wall, significantly reducing <jats:italic>C. albicans<\/jats:italic> viability in cultures and complete inhibition in oral cancer patient samples. Beyond biomedical applications of antifungal drugs, plant cell\u2010based enzyme market is anticipated to double from $52 to &gt;$100 billion in 2028. Edible leaf\u2010based enzyme production offers a new platform that is different from seed\u2010based current technologies for various biotechnology applications.<\/jats:p>","DOI":"10.1111\/pbi.70046","type":"journal-article","created":{"date-parts":[[2025,5,27]],"date-time":"2025-05-27T09:40:56Z","timestamp":1748338856000},"page":"2689-2703","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Expression of mannanase and glucanases in lettuce chloroplasts and functional evaluation of enzyme cocktail against <i>Candida albicans<\/i> in oral cancer patient samples"],"prefix":"10.1111","volume":"23","author":[{"given":"Iqra","family":"Fatima","sequence":"first","affiliation":[{"name":"Department of Basic &amp; Translational Sciences, School of Dental Medicine University of Pennsylvania  Philadelphia PA 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