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This study was aimed to assess the effect of membrane structure on the stability of peanut oil bodies extracted by enzyme-assisted extraction. The influence of pH, NaCl concentration, and temperature on the physicochemical properties of peanut oil bodies was characterized using ζ-potential and particle size. The results indicated that the peanut oil bodies had strong stability (ζ-potential, >20 mV) at pH values away from the isoelectric point (pH 4.8), at a low salt concentration (NaCl concentration, <10 mM), and in a certain temperature range (35 to 55 °C). The stable structure of the oil body was closely related to its structure. Phospholipids, along with membrane proteins, were major components of the oil body membrane. Therefore, the phospholipid composition and content were measured and the types of membrane proteins of the oil bodies were identified. The results showed that phosphatidylcholine and phosphatidylserine were major components of the oil body phospholipids. Two-dimensional electrophoresis showed that the oil bodies contained both intrinsic proteins and extrinsic proteins, which might play an important role in the stability of oil bodies during enzyme-assisted extraction processing.