Extraction Method
To remove any dust particles repeatedly, The pineapples were washed, peeled, and rinsed with tap water and distilled water. Then they were cut into small pieces and blended using a fruit juice processor with a ratio of pineapple to purified water 1: 1.5. It filtered with Whatman number 01 filter paper and obtained supernatant filtrate which was used for further studies.
Sample Purification and Dried
Bromelain enzyme was purified using (NH4)2SO4 concentration of 10%, 15%, 20%, 25%, and 30%, then precipitate it for 12 hrs and 5% ethanol was added. The solution was separated using membrane ultrafiltration. For 12 hrs at 4 o C Bromelain enzyme was precipitated. the clear supernatant collected is centrifuged and then dried by vacuum dried at 45 o C.
Bromelain Activity
By Qualitative analysis of the bromelain, Determination of the bromelain activity is carried out. By knowing the enzyme activity in the bromelain enzyme powder, the performance of the bromelain enzyme can be seen. Bromelain activity of 1 unit means the amount of bromelain required to produce 1 μ mol of tyrosine after 1 minute of digestion of a standard casein substrate at 37 ° C. Specific activity is the ratio of the no. of enzyme activity units of a particular fraction to the protein concentration. The specific activity was used as an index of the purity of the enzyme fraction; the purer the enzyme when the specific activity is High. The ratio of the specific activity of the enzyme fraction to the specific activity of the initial fraction (crude extract) is known as the Purification factor. how effective each refining step is it measures. The ratio of the enzyme units retained after that step to the enzyme units present in the initial fraction (crude extract) expressed as a percentage is the result of the purification step. How well enzyme activity has been preserved or maintained this result can be measured. The enzyme activity was found from the optimization studies to be higher in precipitate with 5% ethanol and 15% ammonium sulfate and using membranes
Bradford Assay:
Bovine serum albumin (BSA) standard (800 ߤL) or the test sample solution was pipetted into a dry test tube. Bradford reagent was added to each tube and mixed thoroughly. At room temperature, the samples were incubated for 10 minutes. The absorbance of the samples was measured using a UV-visible spectrophotometer at a setting of 595 nm.
Benefits:
- Bromelain is used as a dietary supplement for reducing pain and swelling, especially of the nose and sinuses, gums, and other body parts after surgery or injury.
- It is used for osteoarthritis, cancer, digestive problems, and muscle soreness.
- Topical bromelain is used for burns.
Drug Interaction:
Some antibiotics in humans, Bromelain have been documented to increase blood and urine levels, Because of its anti-platelet aggregation and fibrinolytic effects, it may theoretically potentiate the effect of blood thinners. It has historically been used pre-surgery to speed healing.
Dosage:
Bromelain has demonstrated therapeutic benefits in doses as small as 160 mg per day; however, it is thought for most conditions, at doses of 750-1,000 mg daily the best results occur. Most research on bromelain has been performed utilizing four divided daily doses that results are generally dose-dependent.
Side effect and Toxicity:
Bromelain can cause vomiting, rapid heartbeat, stomach upset, diarrhea, and heavy menstrual periods. It can trigger reactions in people with allergies to pineapples, certain pollens, carrots, wheat flour, bee venom, latex, other substances, etc.
Reference:
Maurer HR. Bromelain: biochemistry, pharmacology, and medical use. Cellular and Molecular Life Science; IP; Ahle NW, Hamlet MP. Enzymatic frostbite eschar debridement by bromelain. Ann Emerg Med. 1987; 16(9): 1063-1065.
