EXTRACTION, PURIFICATION OF BROMELAIN FROM PINEAPPLE AND DETERMINATION OF ITS EFFECT ON BACTERIA CAUSING PERIODONTITIS

The pineapple (Ananas comosus) is a multiple fruit belonging to the Bromeliaceae family, has a crude extract   Bromelain accumulated in the stem, fruit, peel and leaves of the pineapple which shows proteolytic activity. The present study was carried out to assess the accumulation of bromelain in the different parts of the pineapple. The techniques homogenization with sodium acetate buffer, followed by filtration and centrifugation were used to extract bromelain from pineapple. The crude bromelain was then purified by ammonium sulphate salt precipitation, dialysis and finally subjected to ion exchange chromatography. Lowry’s method and gelatine digestion unit (GDU) analytical method were performed to estimate the concentration and activity of the crude enzyme and purified bromelain from the different parts The molecular weight of the purified enzyme was determined by SDS-PAGE (sodium dodecyl sulphate – polyacrylamide gel electrophoresis) analysis and found to be 25 to 27 KDa. The final step was to find out the effect of bromelain from leaves and stem of pineapple on pathogens of periodontitis. The enzyme exhibited proteolytic activity against certain bacteria and its capacity to enhance the activity of antibiotics on pathogens causing periodontal diseases. The results of the study confirmed that the enzyme has a high commercial value and a wide range of applications in the field of medicine and biotechnology