Dioscorea alata L. (Dioscoreaceae), commonly known as water yam, is a popular tuber crop vegetable used across the globe. It is mainly composed of starch (75–84% of the dry weight) with a small amount of proteins, lipids, and vitamins and is very rich in minerals. It has nutritional superiority compared with other tropical root crops. The main aim of the study was to extract and characterize the fructooligosaccharides (FOSs) fractions from the tuber of this plant and evaluate their prebiotic properties. The soluble FOSs were fractionated by the conventional solid-phase extraction method and were characterized by FT-IR, HPTLC, and ESI–MS analyses. In addition, the beneficial health attributes of these fractions were checked for in vitro and in vivo prebiotic properties in Swiss albino mice. The FT-IR, HPTLC, and ESI–MS analyses of the FOSs from the hot water and 80% ethanol fractions showed that both are rich in inulin-like prebiotics. Furthermore, these FOSs showed an excellent prebiotic activity score and had enhanced in vitro cholesterol-binding activities in the presence of probiotic lactic acid bacteria (LAB) strains. Moreover, these had good antioxidant efficacies with IC50 values of 36.51 µg/ml and 33.12 µg/ml. In vivo treatment in Swiss albino mice, these FOSs had significantly lowered serum cholesterol (df = 6, p value < 2.2e−16), serum HDL (df = 6, p value = 3.68e−11), serum LDL (df = 6, p value < 2e−16), serum VLDL (df = 6, p value = 2.2e−13), serum triglycerides (df = 6, p value < 2e−16), blood sugar (df = 6, p value = 0.01857), serum glutamic pyruvic transaminase (S.G.P.T., df = 6, p value = 0.01189), serum glutamic oxaloacetic transaminase (S.G.O.T., df = 6, p value = 0.01239) but no significant difference found in weight gain (df = 6, p value = 0.166) and serum creatinine (df = 6, p value = 0.987). Furthermore, they enhanced IgA-mediated immunomodulation and influenced the gut colonization of probiotic LAB strains than the standard prebiotics, inulin. Thus, the study revealed that the FOSs from the storage tuber of D. alata possess some novel prebiotic that could contribute to human nutrition. This is the first report of the prebiotic FOS characterization from this plant.
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