The study attempts the optimization of the total flavonoid content (TFC) and the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) antioxidant activity of the white radish (Raphanus sativus L.) root ethanolic extract (WRE) with regard to several parameters including ethanol concentration, the ratio of solvent/material, temperature and extraction time. Then antifungal analysis of WRE was performed against four fungal species including Aspergillus flavus NBRC 33021, Aspergillus niger NBRC 4066, Aspergillus clavatus NBRC 33020, and Fusarium solani NBRC 31094. At the WRE concentration of 75 mg/mL, diameters of inhibition zone were 9.11 ± 1.5, 19.55 ± 1.68, 17.72 ± 0.25, and 17.50 ± 0.73 mm respectively against the four examined species. Minimum fungal concentration (MFC) values of WRE against the four species were 30, 10, 20 and 30 mg/mL respectively. Evaluation of the sponge cake added with white radish root extract suggested that shelf-life of the sponge cake is 8 weeks in air-cooled condition (20 °C) and two weeks in warm condition (37 °C).