Mycoprotein Production from Mixed Cultures of Rhizopus oligosporus and Neurospora intermedia : Nutritional and Biological Activity for Sustainable Biomass

Abstract The increasing global demand for protein has prompted research into filamentous fungi-based single-cell proteins, particularly regarding their potential contributions to food security and sustainable agriculture. Mycoprotein, generated through the fermentation of filamentous fungi, is characterized by high protein content and notable nutritional and biofunctional properties. This study investigated the nutritional composition and biological activity of a novel mycoprotein produced by coculturing Rhizopus oligosporus InaCC F74 and Neurospora intermedia InaCC F226, utilizing glucose and potato peel residues as substrates. Data were analyzed using ANOVA to assess the effectiveness of the intervention and to determine statistical significance between groups. Mixed culture fermentation was found to enhance the nutritional, physicochemical, and biofunctional properties of mycoprotein relative to monoculture. In the optimized sample, biomass, moisture, protein, lipid, carbohydrate, and ash in optimized sample were 5.62±0.82 g/L, 71.03±1.54, 20.43%, 2.01±0.28, 10.48±1.09%, and 0.59±0.01, respectively. The mixed culture produced a total phenolic content of 2.64±0.19 mg GAE/g DW and a flavonoid content of 2.01±0.14 mg QE/g DW. FTIR analysis indicated the presence of more complex polysaccharide cell wall structures in the mycoprotein. The mixed culture also demonstrated antioxidant and antidiabetic activities, with half maximal inhibitory concentrations (IC50) of 117.41±0.90 mg/mL (DPPH), 13.74±1.78 mg/mL (ABTS), and 2578.08 mg/mL for anti-alpha-amylase activity. Collectively, the mixed culture exhibited superior properties compared to monocultures, with significant improvements observed across all measured parameters.