This study investigated the chemical composition, in vitro cytotoxicity, anti-mildew, and anti-wood-decay fungal activities of the essential oil isolated from the fruit of Liquidambar formosana from Taiwan. The essential oil from the fresh fruit was isolated using hydrodistillation in a Clevenger-type apparatus, and characterized by GC-FID and GC-MS. A total of 45 compounds were identified, representing 98.5% of the essential oil. The main components identified were α-pinene (16.8%), β-caryophyllene (10.1%), -muurolol (8.3%), -cadinol (7.6%), β-pinene (6.7%), and sabinene (5.7%). The essential oil exhibited cytotoxic activity against human oral, liver, and lung cancer cells. The active source compounds were β-caryophyllene, -cadinol, and -muurolol. The fruit essential oil was shown to have excellent anti-mildew and anti-wood-decay fungal activities, the active compounds being evaluated as -cadinol and -muurolol.