Bromelain is a cysteine protease which is found in the tissues of Bromeliacea plant family of which pineapple Ananas cosmossus is best known. The investigated parameters for optimal Brome-lain extraction are optimum buffers, pH, Molarity, time, and amounts of husk free pineapple fruit to volume (ml) of buffer ratio. Sodium phosphate was best buffer for bromelain extraction from pine-apple fruit because its showed high activity, with casein as a substrate. Subsequent experiments, using sodium phosphate as an optimal buffer for extraction and casein as a substrate, reviled that the optimal bromelain extraction conditions were achieved at pH 7.0, 0.1 M of sodium phosphate, 2.5 min of extraction time, 1:0.5 (gm of pineapple fruit/ v of sodium phosphate buffer) extraction per-centage, and 30 min of incubation time. Additionally, Bromelain extracted from pineapple fruit showed a maximum enzyme activity at pH 7and at 30 min of incubation with casein as substrate.