Raspberry ketone (RK) is a high-value bioactive component in the growing demand for natural compounds in food and flavour industries. However, limited production persists due to low-yield microorganisms and complex biosynthetic pathway. Here, an Nidula niveo-tomentosa fungi-based fermentation system was developed to investigate and optimize the cultivation parameters for natural RK production. While optimizing glucose and phenylalanine concentrations ensured sufficient substrate supply, it unexpectedly induced stress on the substrates. Despite having minimal impact on biomass growth, UV-A radiation effectively boosted RK biosynthesis, but also reaching a ceiling with extended exposure times. Younger precultures exhibited higher specific productivity, older cultures ultimately yielded more RK due to their higher inoculum concentration. Mycelial fungi, compared to fungal pellets, outperformed in RK production, likely facilitated by enhanced nutrient uptake and bioproduct transport. Ultimately, the combination of optimized parameters unlocked a peak RK yield of 154.4 mg/L, underscoring the remarkable potential for natural RK production.