AbstractAqueous enzymatic extraction (AEE) of oil from Acer truncatum Bunge seed kernel was investigated. The effects of enzyme type on the extraction yield of oil were studied, and the results showed that the oil yield obtained with pentosanase was higher than that obtained with the other enzymes. The combination of pentosanase and cellulase showed better extraction performance than a single enzyme, and the operation parameters of the AEE method were optimized. A maximum oil yield of 37.94% was obtained. The analysis results of chemical compositions of the extracted oils showed that the content of unsaturated fatty acids in the oil extracted by the AEE method was 90.28%, and the content of nervonic acid was about 5.59%. In addition, the main physical and chemical properties of A. truncatum Bunge seed oil were measured. The oil obtained by the AEE method met the China National Standard of A. truncatum Bunge oil.