Isolation And Analysis Of Intracellular Active Substance From Spirulina Maxima

The hot water-soluble intracellular polysaccharides(IPS), were isolated from Spirulina maxima powder, and deposited by ethanol, and dehydrated by acetone and aether, and decolorized by active carbon, dried and weighed, and 9.8% rough IPS were extracted from Spirulina maxima. PH of the IPS solution was adjusted to PI of protein so that protein deposited and was removed, and then trypsin was put into IPS solution to decompose protein, and then the amino acides from the protein of IPS solution were removed by dialysis. Two fractions, IPS A and IPS B was obtained by Column chromatography of IPS on DEAE-cellulose 52 and Sephadex G-100. Both IPS A and IPS B were homogenous when analyzed by UV-spectrophotometer and Sephadex G-200 column chromatography. Their primary structures were analyzed by a combination of HPLC and IR. IPS A with 238KD relative molecule mass, was mainly composed of L-fucose, L-rhamnose, D-xylose and D- mannose (the molar ratio, 1.0: 0.62:0.99:0.013), uronic acids(13.4μg/mg) and sulfate(26.3μg/mg). IPS B with 34KD relative molecule mass, was mainly composed of D-xylose, L-rhamnose, D-fructose , D-glucose., L-focose and D- galactose (the molar ra.tio, 0.011:0.84:1.0:0.051:0.026:0.0043) and sulfate (21.5μg/mg).A glycoprotein was also isolated from Spirulina maxima powder. Purification of this glycoprotein was accomplished by gel filtration on Sephadex G-200 and SDS-PAGE. The relative molecule mass of the glycoprotein was 136KD. Characterization of this glycoprotein by IR analysis showed the typical absorption of polysaccharide and indicated the presence of α-glycoside linkage . β -Elimination reaction demonstrated the existence of o-glycosidic linkage in this spirliuna glycoprotein .