PURIFICATION AND CHARACTERIZATION OF RHODANESE FROM THE LEAVE OF BITTER MELON (MOMORDICA CHARANTIA)

The Mormodica charantia leaves used in this work was obtained from Ile-Ife, Osun State. Mormodica charantia rhodanese was purified using 80% ammonium sulphate precipitation and affinity chromatography technique. The enzyme had a specific activity of 5.4 Rhodanese Unit per milligram of protein (RU/mg) with a purification fold of 2.42 and a percentage yield of 8.78%. The Km of rhodanese from Mormodica charantia for sodium thiosulphate (Na2S2O3) and potassium cyanide (KCN) were 16.67mM and 20mM respectively, while their Vmax were 0.2RU/ml/min and 0.24RU/ml/min respectively. The substrate specificity showed that the enzyme was not inhibited by 2-mercaptoethanol, ammonium persulphate and sodium metabis- sulphite. The optimum temperature was 60°C at a pH of 7.0. The enzyme was not inhibited by salts (KCl, NaCl, NiCl2, MnCl2, ZnCl2 and BaCl).