Pineapple (Ananas comosus) is a multiple fruit belonging to the family Bromeliaceae, has a crude extract of bromelain accumulated in the stem, fruits, peel and leaves of the pineapple that shows proteolytic activity. The present study was performed to evaluate the accumulation of bromelain in different parts of the pineapple. Homogenization techniques with sodium acetate buffer, followed by filtration and centrifugation were used to extract pineapple bromelain. The crude bromelain was then purified by precipitation with ammonium sulfate salt, dialysis and finally subjected to ion exchange chromatography. We performed the Lowry analytical method and the gelatin digestion unit (GDU) analytical method to estimate the concentration and activity of crude enzyme and purified bromelain from different parts. The molecular weight of the purified enzyme was determined by SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis) and found to be 25 to 27 KDa. The final step was to discover the effect of leaf bromelain and pineapple stem on periodontitis pathogens. The enzyme showed proteolytic activity against certain bacteria and its ability to enhance the activity of antibiotics in pathogens that cause periodontal diseases.
Bromelain is a major protease enzyme found in the pineapple plant (Ananascomosus). It has been chemically known since 1876 and was first identified by Marcano in 1891. Research and isolation of bromelain has been initiated since 1894. Proteolytic enzymes sulfhydryl are the main constituents of bromelain. Bromelain is abundant in the stem and fruit of the pineapple plant and can also be isolated in a small amount of pineapple residue such as core, leaves, peel, etc ... The bromelain present in the pineapple fruit has assigned EC number EC 3.4.22.33 and is considered as fruit bromelain (FBM). Similarly, the bromelain present in the stem of the pineapple is called stem bromelain (SBM) and its EC number is EC 3.4.22.32. The bromelain of the stem has different biochemical properties and composition in comparison with the bromelain of the fruit and contains a varied mixture of different thiol-endopeptidasas. Researchers are making efforts to achieve highly purified bromelain in fewer steps and at lower cost. Modern strategies such as membrane filtration, reverse micellar systems, two-phase aqueous extraction and chromatographic techniques have shown promise in this regard. The pineapple plant also contains minor amounts of other proteinases such as ananain and comosain but bromelain is considered as a primary and widely investigated component. The reason for being so valuable is due to its miraculous use as a phytomedicine. Bromelain has anti-oedematous, fibrinolytic, anticancer, anti-inflammatory, antibiotic, anticoagulant and antithrombotic functions (Kavitha et al., 2013). It is also a possible postoperative agent that helps healing and reduces post-surgical discomfort and swelling. In addition to the clinical approach, bromelain has also been used in a variety of industries including food industries such as breweries and meat processing and softening, textile and cosmetic industries. With the advent of recombinant DNA technology, scientists and researchers around the world have been working on recombinant bromelain to achieve exaggerated and novel applications in the future.