Haseeb Akram Sindhu, Abu Saeed Hashmi, Faiza Masood and Muhammad Sajid
Institute of Biochemistry and Biotechnology (IBBT), UVAS, Lahore, Pakistan.
Fig (Ficus carica) and Kachri (Cucumis trigonus) contain high concentration of proteolytic enzymes. In current research work, purification and characterization of papain (a proteolytic enzyme) from Kachri (Cucumistrigonus) and Fig (Ficus carica) was carried out. Kachri and Fig was crushed separately in a food blender byusing 0.1M (PO4)-3 buffer of pH 8.00, distilled water and 70% ethanol. Liquid mixture obtained was centrifuged and supernatant used as enzyme source. Purification of papain was carried out by Ammonium Sulphate precipitation and dialysis followed by Gel filtration by Sephadex G-50. Then characterization of papain such as protein estimation, determination of proteolytic activity (International Unit) of enzyme and SDS-PAGE analysis were performed to determined molecular weight. Finally, the yield and proteolytic activity of papain was measured and compared with the commercial products. Crude preparation of enzyme has a wide specificity due to the presence of various proteinase and peptidase isozymes. These enzymes being present in natural fruits were free from any toxic effects and consequently can be used in food and pharmaceutical industries.
Key words: Kachri, Fig, Papain, SDS-PAGE, Sephadex G-50, Isozymes