BACKGROUND: Wines are produced via the alcoholic fermentation of suitable substrates, usually sugar (sugar cane, grapes) and carbohydrates (wheat, grain). However, conventional alcoholic fermentation is limited by the inhibition of yeast by ethanol produced, usually at approximately 13-14%. Aside from that, soursop fruit is a very nutritious fruit, although it is highly perishable, and thus produces a lot of wastage. Therefore, the present study aimed to produce fermented soursop juice (soursop wine), using combination of two starter cultures, namely mushroom (Pleurotus pulmonarius) and yeast (Saccharomyces cerevisiae), as well as to determine the effects of fermentation on the physicochemical and antioxidant activities of fermented soursop juice. Optimisation of four factors (pH, temperature, time and culture ratio) using response surface methodology were performed to maximise ethanol production.RESULTS: The optimised values for alcoholic fermentation were pH 4.99, 28.29 °C, 131 h and a 0.42 culture ratio (42:58, P. pulmonarius mycelia:S. cerevisiae) with a predicted ethanol concentration of 22.25%. Through a verification test, soursop wine with 22.29 ± 0.52% ethanol was produced. The antioxidant activities (1,1-diphenyl-2-picrylhydrazyl and ferric reducing antioxidant power) showed a significant (P