This study investigated the effects of enzymatic treatment of peanut butter on two-major peanut allergens (Ara h 1 and Ara h 2). Home-made and commercial peanut butter samples were treated with alpha-chymotrypsin, trypsin or the combination of these enzymes and incubated at room temperature for 24 h or at 37 °C for 3 h. Treated peanut butter samples were sampled weekly for evaluation of total soluble proteins and extractable Ara h 1/Ara h 2. Data show that 1:1 alpha-chymotrypsin: trypsin at 0.04% of enzyme-to-peanut butter ratio resulted in near complete reduction of extractable Ara h 1 and Ara h 2 respectively. Treatment of peanut butter with a combination of trypsin and alpha-chymotrypsin resulted in a decrease in IgE-binding, suggesting that enzymatic treatment has the potential to reduce the allergenicity. However, clinical tests are needed to confirm any reduction in allergenic potential. The amount of water used to disperse enzyme did not have significant effect on allergen reduction but affected the consistency and colour of treated products, especially when the amount of water added was above 5% of peanut butter weight.