Technical Abstract: This study investigated the use of enzymatic treatment to reduce peanut allergens in peanut kernel by processing conditions, such as, pretreatment with heat and proteolysis at different enzyme concentrations and treatment times. Two major peanut allergens, Ara h 1 and Ara h 2, were used as indicators of process effectiveness. The data show that the proteases used in this study were very effective in reducing Ara h 1 and Ara h 2 in roasted peanut kernels, but less effective when used in raw peanut kernels. Treatment of roasted peanut kernels using solutions of chymotrypsin and trypsin for 1-3 hr significantly increased the solubility of peanut protein, and reduced the Ara h 1 and Ara h 2 in peanut kernel extracts by as much as 90-100% based on ELISA readings. The Ara h 1 and Ara h 2 contents in peanut protein extracts were inversely correlated to the solubility of roasted peanut protein (R2=0.8-0.99). Blanching of kernels enhanced the effectiveness of enzyme treatment in roasted peanuts but not in raw peanuts. The optimal concentration of enzyme solution was determined to be in the range of 0.1 to 0.2%. At low enzyme concentration (0.01%), treatment time has significant effect on allergen reduction; however, time became less significant at enzyme concentration of 0.05% or higher. At the optimal enzyme concentration range; the process was fast, yielding 99% reduction of the extractible Ara h 1 and Ara h 2 after just 15 minutes of incubation. Treatment of peanut kernels by proteases may be an effective way to produce peanuts with higher soluble material, but degraded allergens relative to untreated peanuts.