Extraction of papain enzyme from papaya leaves using enzyme assisted method

Papain is highly appreciated in medical field that prevents several chronic diseases such as cardiovascular disease, cancer and diabetes. Therefore, the objective of the research is to extract papain enzyme from papaya leaves using enzyme assisted method and determine papain activity. The Cellullase assisted extraction process was optimized by varying different parameters such as pH of extraction process, Cellulase concentration, Solid to Liquid ratio, incubation time and incubation temperature. Each 5 g of papaya leaves were ground and mixed with acetate buffer at a different Solid to Liquid ratio (1:5 - 1:25 g/ml) and adjusted with different pH (3-8). Cellulase was quantified and dispersed in acetate buffer (Na2HPO—citric acid) in different concentration (0.5 - 2.5 mg/ml).The enzymatic pretreatment was carried out and continued for enzyme assisted extraction process for various temperature and time (5070o4C and 1-5 h respectively). The optimal extraction conditions that satisfied the above constraints were found to be at pH 7, 1.5 mg/ml of Cellulase, solid to liquid of 1:10 (g/ml), an extraction time of 4 h and at an extraction temperature of 65oC. Under the optimum conditions, the extraction yield of papain was successfully achieved as much as 3.8018 µmole with 2.0910 Units/ml in Cellulase treated sample. Therefore, the potential for the papain enzyme in medical field can be fulfilled using enzyme assisted extraction.