Endophytic fungi isolated from plants of different ecosystems have an impressive ability to produce diverse chemical classes of bioactive secondary metabolites. In this study, the cytotoxic activity and chemical composition of the ethyl acetate extract of the white beans culture of the endophytic fungus (FWBC), Talaromyces wortmannii isolated from the widespread reed grass, Arundo donax L. were investigated. A new ceramide named talaramide (1), along with other four known compounds; stigmasterol (2), thymine (3), uracil (4), and stigmasterol-3-O-β-D-glucoside (5) were identified. The structure of compound 1 was elucidated based on extensive 1D and 2D NMR spectroscopic techniques in combination with HR-ESI-MS. The produced FWBC extract exhibited cytotoxic activity against three cancer cell lines viz., MCF7, HepG2, and A549 (IC50; 54.57 ±0.27, 57.12 ± 0.59 and 84.33 ± 1.07 µg/mL, respectively) compared to methotrexate (IC50; 80.25 ± 1.11, 83.83 ± 0.74 and 153.20 ± 2.03 µg/mL, respectively) as a positive control. It showed selectivity indices with values ranging from 1.37 to 2.11 using the normal cell line, WI38 (IC50; 115.43 ± 1.92 µg/mL). The observed cytotoxicity, combined with acceptable selectivity indices, suggests the potential of this endophyte as a source of anticancer agents. Based on previous reports, it was also suggested that the identified ceramide could be responsible for the observed cytotoxic activity of the investigated extract.