The reactions leading to the formation of precursors of chocolate flavor are performed by endogenous enzymes present in the cocoa seed. Polyphenol oxidase (PPD) presence and activity during fermentation of cocoa beans is responsible for the development of flavor precursors and is also implicated in the reduction of bitterness and astringency. However, the reliability of cocoa enzyme activities is complicated due to variations in different genotypes, geographical origins and methods of fermentation. In addition, there is still a lack of systematic studies comparing different cocoa cultivars. So, the present study was designed to characterize the activity of PPD in the pulp and seeds of two cocoa cultivars, PH 16 and TSH 1188. The PPD activity was determined spectrophotometrically and characterized as the optimal substrate concentration, pH and temperature and the results were correlated with the conditions during the fermentation process. The results showed the specificity and differences between the two cocoa cultivars and between the pulp and seeds of each cultivar. It is suggested that specific criteria must be adopted for each cultivar, based on the optimal PPD parameters, to prolong the period of maximum PPD activity during fermentation, contributing to the improvement of the quality of cocoa beans.Keywords: cocoa; fermentation; enzyme activity; chocolate.Practical Application: To obtain knowledge for future technological interventions during fermentation for the improvement of the quality of raw material in the production of monovarietal chocolates.