Purpose. Potato fruit juice, a residue of starch industry, contains up to 2.5% [w/w] of proteins that are potentially valuable raw-materials of food, cosmetic, and pharma industries. The recovery of protein from the potato fruit juice is limited by the lack of industrially feasible concentration and separation technologies. The present research thus aimed at development of such process for the separation of active protease inhibitors from potato fruit juice. Methods. Low temperature mechanical vapor recompression evaporation was applied for concentration of potato fruit juice followed by ethanol precipitation for recovery of active proteins. The effects of precipitation temperature and precipitative agents were investigated employing response surface modeling methodology. Results. Concentration of potato fruit juice by evaporation was successful without loss of trypsin inhibition activity. Precipitation using 6.5 M ethanol at low temperature (0-+4°C) was found suitable for the recovery of active protease inhibitors from the concentrate. Piloting at starch industry yielded 50% of total proteins, with a high quantity of active protease inhibitors and a minor inclusion of other proteins. Conclusion. Concentration by low-temperature evaporation, followed by ethanol precipitation of protease inhibitors at optimized temperature, is an attractive option for valorization of potato fruit juice.