Characterization of Metabolites in Plant-Based Milk Yogurt Enriched with Wolffia globosa to Improve Bionutritional and Functional Properties

Riceberry rice milk (RBRM) is rich in phytochemicals, particularly anthocyanins, which are known for their potential in managing type 2 diabetes (T2D). This study aimed to develop a novel RBRM-based yogurt derived from its polysaccharide and protein components and to evaluate the effects of supplementation with W. globosa powder (WGP) at 0% (F1, control), 5% (F2), 10% (F3), and 15% (F4) on nutritional and functional properties. Among all formulations, F4 exhibited the highest nutritional values, including dietary fiber (41.25%), curd protein (21.34%), and carbohydrate (starch) content (25.25%), with a lower fat content (2.13%) compared to other groups. In terms of antioxidant activity, F4 showed high total phenolic content (33.70 mg GAE/g) and total flavonoid content (25.2 mg QUE/g), along with strong radical scavenging activities, with DPPH and ABTS inhibition values of 41.52% and 78.18%, respectively. Furthermore, F4 demonstrated notable antidiabetic potential through α-amylase and α-glucosidase inhibition, with IC50 values of 0.89 and 1.32 mg/mL, respectively. Widely targeted metabolomics analysis identified 88 differential metabolites between F4 (potent condition) and F1 (control group). Twelve selected compounds from RBRM–WGP yogurt contributed to increased levels of amino acids, peptide derivatives, saccharides, organic acids, polyphenols, and flavonoids. Molecular docking analysis revealed that key metabolites, including vignatic acid B, glimepiride, and indoramin, exhibited strong binding affinities with the active sites of α-amylase (PDB: 2GVY, Aspergillus niger) and α-glucosidase (PDB: 3A4A, Saccharomyces cerevisiae). These findings indicate that phytonutrient compounds, particularly indoramin, play a significant role in enhancing the nutritional composition and functional properties of RBRM–WGP yogurt for potential applications in food processing.