This study aimed to investigated the effect of adding increased amounts of banana flour (0, 0.5, 1.5 e 3 w/w %, I, II, III, IV) in probiotic fermented milk processing. Microbiological (Streptococcus thermophilus, Lactobacillus acidophilus and Bifidobacterium counts), Physico-chemical analysis (gross composition, resistant starch, pH, proteolysis and firmess) were performed weekly along 21 d. While S. thermophilus presented always values about 8 log cfu/g for all samples, probiotic microrganisms presented different behavior. While L. acidophilus presented a decrease in the viable counts in the samples added with 3 w/w % banana flour (from 8.03, d 1, to 7.14 log cfu/g, d 21, P < 0.05), Bifidobacterium present increase along the storage time (6.78, d 1 to 7.14 log cfu/g, d 21, P < 0.05). Toward the physic-chemical parameter, pH did not present difference among the samples and related to storage time (values ranged 4.60 to 4.67, P > 0.05), while increased proteolysis level was observed at the fermented milks added with intermediate levels of banana flour (values ranged from 0.430 to 0.667, sample II and 0.455 to 0.662, sample III) in relation to sample III (values ranged from 0.345 to 0.378, P < 0.05). For the firmness, it was observed direct relation with the banana flour addition (values ranged to 1.23, sample I to 1.88 N, sample IV) without relation to the storage time. The gross composition reported absence of difference among the fat and proteins levels while for resistant starch content, sample III presented increased decrease (30.11%) compared with sample IV (20.07%), respectively. Our findings suggest the need to adjust the banana flour dosage during probiotic fermented milk processing as well as the probiotic culture to be added at the formulation to obtain the prebiotic effect.