In the present study, a prebiotic acerola juice containing gluco-oligosaccharides and dextran was produced and processed by high-intensity ultrasound (HIUS). After a simulated in vitro digestion, the glucooligosaccharides and dextran maintained 90% and 80% of their initial concentration in all prebiotic's juices. At the same time, Vitamin C and phenolic compounds concentration increased significantly by 19% and 7% (P < 0.05). After the in vitro digestion, the prebiotic juice HIUS processed by 10 min showed the highest increase in gluco-oligosaccharides and bioactive compound concentrations. The HIUS processing imparted some dextran hydrolysis and improved its fermentability by Lacticaseibacillus casei. Gluco-oligosaccharides were extensively consumed as substrate in simulated intestinal conditions, promoting the L. casei NRRL B-442 growth and production of organic acids and short-chain organic acids. The prebiotic juice HIUS processed for 6 min showed the best responses regarding the metabolism of L. casei NRRL B-442. The results showed high-intensity ultrasound processed acerola juices containing glucooligosaccharides and dextran as a prebiotic food.