Extraction of soybean oil from flaked and extruded soybeans using enzyme-assisted aqueous extraction processing (EAEP) is a promising alternative to conventional hexane extraction. The efficiencies of four proteases releasing oil from extruded material were compared. Protex 51 FP, Protex 6L and Protex 7L each extracted 90% of the total oil available while Protex 50FP gave similar extraction yield as the control (without enzyme treatment). During EAEP, however, a stable emulsion forms that must be broken in order to recover free soybean oil. The potential of various proteases and phospholipases to destabilize the emulsion was determined. Two enzymes, a phospholipase A2 (LysoMax(TM)) and a protease (Protex 51 FP) were selected to determine the effect of enzyme concentration on demulsification. Although at a 2% concentration (w/w, enzyme/(cream + free oil)), each enzyme tested was effective in totally destabilizing the cream; the protease released significantly more free oil than did the phospholipase at concentrations less than 2%. At 0.2% concentration, 88 and 48% of free oil were obtained with the protease and phospholipase, respectively. Reducing the pH of the cream also destabilized the cream with maximum demulsification at the isoelectric point of soy proteins. These results provide destabilization strategies for the oil-rich emulsion formed during aqueous extraction processing of extruded flakes and significantly contribute to the development of this environmentally-friendly technology. Published by Elsevier Ltd.