Extraction of Bromelain and Investigation of Proteolytic Ability of the Stem and Fruit in Three Different Pineapple Cultivar of Morris Pineapple (Ananas comosus L. var. comosus Morris), Josepin Pineapple (Ananas comosus L. var. comosus cv. Josephine), and MD2 Pineapple (Ananas comosus L. Merrill cv. MD2).

Lim, Yi Xiang (2018) Extraction of Bromelain and Investigation of Proteolytic Ability of the Stem and Fruit in Three Different Pineapple Cultivar of Morris Pineapple (Ananas comosus L. var. comosus Morris), Josepin Pineapple (Ananas comosus L. var. comosus cv. Josephine), and MD2 Pineapple (Ananas comosus L. Merrill cv. MD2). Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Bromelain is a proteolytic enzyme which is referring to either of two protease enzymes that can be found in the plants under Bromeliaceae family. These two proteases enzymes are Fruit Bromelain that can be found in the fruit and the peels of the pineapple and also Stem Bromelain that can be found in the stem of pineapple plant. The objectives of this experiment are to determine the best source of Bromelain among different parts and different cultivar of pineapple, to test the proteolytic ability of the extracted Bromelain and also to fully utilize the pineapple waste. In this experiment, crude Bromelain was extracted from different parts (fruit and stem) of different cultivar, Morris pineapple (Ananas comosus L. var. comosus Morris), Josepine pineapple (Ananas comosus L. var. comosus Josapine) and MD2 (Ananas comosus L. Merrill cv. MD2). The crude Bromelain extracted will be blended with sodium phosphate buffer then centrifuged. The supernatant will undergo salting precipitation and dialysis before analysis was carried on. Concentration of bromelain extracted from different parts of different cultivar of pineapples can be calculated by using Lowry’s method and known concentration BSA solution was used as standard curve. Bromelain extracted from Josepine stem shown highest concentration among three cultivars which is 131 ± 60ppm per 100ml of extracted Bromelain solution. The proteolytic ability of Bromelain extract was also tested by using casein as substrate. Casein was hydrolyzed by bromelain and free tyrosine was released by the casein. Amount of tyrosine released can be calculated by measuring absorbance using spectrophotometer and known concentration of tyrosine solution was used to plot the standard curve. In the same volume of Bromelain solution, Bromelain extracted from Josapine pineapple’s fruit shows the highest proteolytic ability among three cultivars where 7.865ppm of tyrosine is release per 5ppm if Bromelain solution used. Josapine stem is the best source for the extraction of Bromelain as stem is cheap, easily obtainable and highest concentration of Bromelain can be extracted. However, Josapine Fruit Bromelain shows the strongest proteolytic ability among all samples tested. Key word: Josepine Pineapple, Morris Pineapple, MD2 Pineapple, Bromelain, Lowry’s method, Enzymatic hydrolysis of casein.