| The aim of this study was to apply first and second dimension Electrophoresis together with zymogragraphy technique to analyse the presence of hemolytic activity of the earthworm (Lumbricus rubellus) extract. Earthworms were obtained from Pusat Studi Satwa Primata (PSSP), Bogor. Extraction was conducted by verying the pHs from pH 9 to pH 3 and vice versa. Sample was also been purified using AKTAprime desalting column with Sephadex G-25. The results showed that there were 12 spots of hemolysin protein in 2 D zymogram, which was separated further from the 9 fractions obtained from the one dimension Zymography. The smallest molecule appeared about 15 kD and the biggest appeared about 37 kD with unsignificant molecular weight difference. And the isoelectric points were at pH 4,6-5.1. Comparison of the 2D Electrophoresis and Zymography revealed similar and different spots which imply that some protein molecules fractions possess both fibrinolytic, hemolytic activity and there could be another activity. The result also showed that the enzyme derived from earthworm has a great capability to refold after being treated with high voltage, urea, Dithiothreitol (DTT), and Iodoacetamide applied in 2 D electrophoresis procedure. |