Fruit smoothies are nonalcoholic beverages perceived as healthy and refreshing beverages well-accepted by consumers of all ages. These products have been considered suitable matrices for incorporation of probiotic cultures because can be consumed by lactose intolerant and vegetarian individuals. The objective of this work was to assess the physicochemical parameters of fruit smoothies added of Lactobacillus acidophilus (LA-03) during refrigerate storage (4 ºC). The smoothies were formulated with açaí (Euterpe oleracea Mart.) and banana (Musa paradisiaca L.) (80:20) or mango (Mangifera indica L.) and passion fruit (Passiflora edulis f. Flavicarpa) (60:40). Each pasteurized smoothie (80 ºC, 10 minutes) was added of the LA-03 culture (1mL/100mL; 8 log CFU/mL) and stored 21 days. On the day 0 (just after the manufacture), 1, 7, 14 and 21 of storage, viable counts (VC) of LA-03, soluble solids (SST), pH, total titratable acidity (TTA), color, and apparent viscosity were assessed. Sugars (glucose and fructose) and organic acids (lactic, malic, and succinic) were also determined. Statistical analysis considered p<0.05. The microbiological parameters (total and thermotolerant coliforms, coagulase-positive Staphylococcus aureus, Salmonella spp. and Listeria spp.) followed the current sanitary standards established by the Brazilian Legislation during the period evaluated. In both açaí- and mango-based smoothies, VC of LA-03 were >7 log CFU/mL over time assessed. After the day 14, the pH of both smoothies decreased p<0.05 (açaí- 4.63-3.95; mango-based 3.91-3.44), while the TTA (açaí- 1.7-2.7; mango-based 8.3-8.8) increased (p<0.05). Apparent viscosity decreased (p<0.05) and Luminosity (L*) (açaí- 60.82-59.95; mango-based 30.68-29.70) did not change (p>0.05) in both smoothies during the time assessed. The changes (p<0.05) in contents of each organic acid or sugar evaluated during the storage intervals varied depending on the smoothie. The results suggest the evaluated açaí- and mango-based smoothies as interesting matrices for addition of the LA-03 culture on development of potentially probiotic smoothies.