PURIFICATION OF HEMOCYANIN FROM MARINE GASTROPOD RAPANA THOMASIANA USING AMMONIUM SULFATE PRECIPITATION METHOD

摘要

The aim of this study was to develop an efficient and simple method for isolation of preparative amounts of pure hemocyanin (He) from the hemolymph of the marine gastropod Rapana thomasiana. The methods that are usually used, as ultracentrifugation and column chromatography, are expensive and impractical for the large-scale production of He. For the ammonium sulfate precipitation method, the concentrated hemolymph was twice precipitated by 38% saturation with crystalline ammonium sulfate. Rapana thomasiana hemocyanin (RtH) was isolated with good yield and high purity, as assessed by gel chromatography, SDS-PAGE, transmission electron microscopy and absorption spectroscopy. This suggests that the ammonium sulfate precipitation is an efficient and useful purification method, suitable for a large-scale RtH preparation.