Bromelain is a general term for proteolytic enzymes containing sulfhydryl groups at the active site, derived from pineapple (Ananas comosus (L.) Merr.), which function in the hydrolysis of proteins. This study aimed to compare the activities of crude bromelain extracts obtained from the fruit, core, and peel of the A. comosus (L.) Merr. cv. Phuket, as well as to investigate the differences between unripe and ripe pineapples. The activity of crude bromelain was determined by measuring the amount of tyrosine released, compared with a tyrosine standard, and expressed in units per milliliter of enzyme. The results revealed that the highest enzyme activity was observed in the extract from the ripe fruit flesh under optimal conditions: pH 4.5, 50 °C and 10 minutes of incubation when using casein as a substrate, which yielded enzyme activity of 5.4 and 5.7 units per milliliter, respectively. Additionally, the efficiency of the crude enzyme extract at pH 4.5 and incubated at 50 °C was evaluated for removing soy sauce stains from cotton fabric. It was found that crude bromelain effectively removed stains, with the highest efficiency observed after soaking the fabric in the enzyme extract for 24 hours. These findings indicate that bromelain from the fruit flesh of A. comosus (L.) Merr. cv. Phuket has high potential for application in the detergent industry and could be further developed into enzyme-based textile care products in the future.