Background: Polysaccharides from the medicinal mushroom Phellinus rimosus (Berk.) Pilát (PR) are the major functional bioactive ingredients. However, there has been a marked natural decrease in the number of PR fruit bodies, leading to their increased cost. Moreover, the natural growth and development of mature PR fruit bodies takes several decades. Objective: The objective of this study was to produce a polysaccharide extract from cultured PR mycelia (PEPRM) by using ultrasonic-assisted extraction with response surface methodology (RSM), and determine its physicochemical composition and antioxidant potential. Methods: Polysaccharide and monosaccharide composition analyses were carried out by Fouriertransform infrared spectroscopy (FT-IR) and High-performance liquid chromatography (HPLC). Total contents of polysaccharides, beta-glucans, phenolic compounds, and flavonoids were investigated utilizing the phenol-sulfuric acid method, enzymatic-based commercial test kit, Folin-Ciocalteu method, and aluminium chloride colorimetric method, respectively. Antioxidant activity was determined by using 2,2-diphenyl-1-picrylhydrazyl-hydrate (DPPH) radical scavenging assay and 2,2- azino-bis (3-ethylbenzothiazol-6-sulfonic acid) (ABTS) radical cation decolorization assay. Results: Optimal conditions for the production of PEPRM included a ratio of 51.29 mL water to 1 g PR mycelia and an extraction time of 46.23 minutes, resulting in a total polysaccharide content of 577.5 mg/g of PEPRM. FT-IR spectra of PEPRM showed two broad bands at 3272.08 cm-1 and 2924.8 cm-1 in the carbohydrate region and the peaks at 1078.44, 1019.05, and 853.0 cm-1 indicated the presence of the pyranose ring skeleton, glycosidic linkage, and glucans. PEPRM had molar ratios of glucose: mannose: rhamnose: fucose, i.e., 21.86: 1.00: 2.08: 3.40, respectively. PEPRM had total contents of beta-glucans, phenolic compounds, and flavonoids as percentages of dry weight, i.e., 21.22, 2.51, and 5.71, respectively. PEPRM showed better inhibitory activity against ABTS radicals than DPPH radicals. result: Optimal conditions for the production of PEPRM were a ratio of 51.29 ml water to 1 g PR mycelia and extraction time of 46.23 min, yielding a total polysaccharide content of 577.5 mg/g of PEPRM. FT-IR spectra of PEPRM showed two broad bands at 3272.08 cm-1 and 2924.8 cm-1 in the carbohydrate region and the peaks at 1078.44, 1019.05, and 853.0 cm-1 indicated the presence of the pyranose ring skeleton, glycosidic linkage, and glucans. PEPRM had molar ratios of glucose: mannose: rhamnose: fucose, i.e., 21.86: 1.00: 2.08: 3.40, respectively. PEPRM had total contents of beta-glucan, phenolic compounds, and flavonoids as percentages of dry weight, i.e., 21.22, 2.51, and 5.71, respectively. PEPRM showed better inhibitory activity against ABTS radicals than DPPH radicals. Conclusion: This is the first finding to reveal that ultrasonic-assisted extraction with RSM was an environmentally friendly alternative to produce antioxidant polysaccharides from cultured PR mycelia. conclusion: This is the first finding to reveal that ultrasonic-assisted extraction with RSM was an environmentally friendly alternative to produce antioxidant polysaccharides from cultured PR mycelia.

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.

This study aimed to illustrate the effects of plant-based yogurts made from bio-fermentative green rice milk yogurt (GRMY), enhanced with Wolffia globosa powder (WGP) and infused with Thai green tea. Different formulations (F1 to F4) were analyzed to assess the impact of varying weight ratios of WGP, ranging from 0% to 15%. The goal was to improve nutritional values and bioactive components. The F4 formulation of plant-based yogurt demonstrated enhanced nutritional values, including higher levels of dietary fiber, starch, protein, and fat contents, which were 42.45, 26.55, 22.48, and 2.29% DW, respectively. It was increased by bioactive compounds such as total phenolic content (52.99 mg GAE/g), total flavonoid content (60.45 mg QUE/g), and β-carotene (1.81 mg/g). Additionally, the F4 formulation exhibited the highest antioxidants in terms of DPPH activity (5.48 mg TROE/g), ABTS activity (424.15 mg TROE/g), and FRAP reducing capacity (106.35 mg TROE/g) and antidiabetic activity of α–glucosidase inhibition in IC₅₀ at 1.41 mg/mL. The yogurts were evaluated through metabolomic network analysis as a complex bioactive food system, wherein fermentation-derived metabolites synergistically interact with tea polyphenols, amino acids, phytosterols, and carotenoids. The prevailing metabolic profiles indicate an increase in antioxidant capacity, a reduction in inflammatory burden, enhanced glucose metabolism, alterations in gut microbiota, cardiovascular protection, and neuroprotective potential. These findings suggest that the product represents a promising plant-based functional food, offering numerous health benefits. This assertion is supported by a diverse metabolite network identified through UPLC–MS/MS metabolomic research.