Michaelis-Menten Parameters Characterization of Commercial Papain Enzyme “Paya”

Abstract

The use of enzymes is increasing nowadays, both for research and for the industry. This happens due to the advances in various fields such as fermentation technology, genetic engineering, and enzyme applications technology. One kind of enzymes that is very widely used and plays an important role in biotechnology and industrial applications is a protease enzyme, especially papain enzyme. Papain is a proteolytic enzyme which is isolated from papaya fruit sap tapping, or it could be derived from the leaves of papaya (Carica papaya L.). Actually, in the market, there is already a commercial papain enzyme called "Paya" which serves as a meat tenderizer with a relatively low price, but the enzyme papain was not pure and has unknown activity. Therefore, this study aims to examine the concentration of dissolved papain enzyme and casein as the substrate. By using the Lowry protein assay method, the initial concentration of papain enzyme and casein respectively 20,000 ppm. The result of protein assay we found that the concentration of dissolved papain enzyme and dissolved casein are respectively 71.069 ppm and 764.8276 ppm. The results from the first experiment are used for the second experiment which is to measure and determine the enzyme kinetics parameters Km and Vmax of that commercial papain enzyme with casein as the substrate. From this research, we found that the Km and Vmax of this commercial papain enzyme respectively 248.68 ppm dan 1.514 ppm casein/minute.