The demand for medicinal mushrooms as dietary supplements is rising due to their health benefits, particularly from secondary metabolites found in their fruiting bodies. Erinacine C, a secondary metabolite synthesized by the edible mushroom Hericium erinaceus, has garnered attention for its potential therapeutic effects, especially in treating Alzheimer's disease. However, current products contain low amounts of erinacine C, and large-scale production through submerged cultivation has not yet been achieved. This study aims to enhance the production of erinacine C through two dosage forms: an edible biomass with increased erinacine C content and purified erinacine C for functional food and pharmaceutical applications. The research explores using biogenic byproducts, such as acidic-treated wheat bran and brewer's spent grain, as precultivation media. Results showed that biomass grown on these byproducts yielded lower erinacine C concentrations. In contrast, using synthetic precultivation media proved more effective, achieving a maximum concentration of 2.7 g/L (300 mg/g dry weight) with optimal conditions. The biomass was extracted with ethyl acetate and purified via centrifugal partition chromatography, isolating 100 mg of erinacine C with over 99% purity. Production costs were estimated at €16,000 per kg for purified erinacine C and €4,000 per kg for food-grade biomass containing 16 wt.-% erinacine C, with biomass costs at €60
