Bromelain is a proteolytic enzyme from Bromeliaceae plants that can be found in pineapple (Ananas comosus (L.) Merryl). Bromelain has activity as a pharmaceutical and food. The aim of this study was to determine the bromelain content from stem bromelain extract (SBM). The stem of pineapple was extracted using a grater and then concentrated by a freeze dryer to obtain a pure concentrate. SDS-PAGE, Bradford assay and protease specific activity was used to analyze bromelain activity. The results of this study showed that bromelain from pineapple stem has a molecular weight between 19-22 kDa. Bradford assay showed 0.476 µg/mL bromelain concentration. Bromelain activity of Non freeze dried (NF) extract showed 0.317 U/mL, while freeze dried (F) extract showed 0.212 U/mL. This study concluded that the pineapple stem has bromelain activity.