Potato protein liquor (PPL), a side stream from the potato starch industry, is normally used as fertilizer. However, with more than 100 g/L of sugars, 20 g/L of Kjeldahl nitrogen and Chemical Oxigen Demand (COD) of 300 g/L, it represents serious environmental challenges. The use of PPL for fungal cultivation is a promising solution to convert this waste into valuable products. In this study, PPL was characterized and used to cultivate edible zygomycete Rhizopus oryzae, which is widely used in Southeast Asian cuisine to prepare e.g., tempeh. Moreover, it can be potentially used as a protein source in animal feed worldwide. Under the best conditions, 65.47 ± 2.91 g of fungal biomass per litre of PPL was obtained in airlift bioreactors. The total Kjeldahl nitrogen content of the biomass was above 70 g/kg dry biomass. The best results showed 51% reduction of COD and 98.7% reduction in the total sugar content of PPL.