PREBIOTIC POTENTIAL OF HYBRID REQUEIJÃO

The partial replacement of animal proteins with vegetable proteins has gained attention in the food industry, and hybrid requeijão with added soy protein is a promising alternative. However, the health effects of this dietary change, particularly on the intestinal microbiota, remain underexplored. This study aimed to evaluate the prebiotic potential of requeijão by partially replacing (50%) dairy protein with soy protein. Control and hybrid requeijões with 12% protein (calcium caseinate or calcium caseinate + soy protein isolate) were produced in a model system. To assess prebiotic potential, the cheeses underwent in vitro gastrointestinal digestion (INFOGEST), and the hydrolysate obtained was tested for its effect on the growth of *Lacticaseibacillus casei*. Growth was monitored by turbidimetry at 660 nm, and standard plate counts measured microbial viability after 0, 24, and 48 hours. Fructooligosaccharide (FOS) served as a prebiotic control. ANOVA was used to evaluate the effects of treatment, incubation time, and their interaction on microorganism growth. Results showed that treatment, incubation time, and their interaction significantly influenced the growth of *L. casei*, with all conditions leading to increased plate counts over time. Hydrolysates from control and hybrid requeijões stimulated the growth of *L. casei* after 24 and 48 hours, with no impact from protein replacement. However, the effect was weaker than that of FOS. These findings suggest that the partial substitution of caseinate with soy protein in requeijão does not hinder the growth of *L. casei*, and both control and hybrid requeijões successfully promoted probiotic growth under the study conditions.