Study On Efficient Extraction And Purification Technology Of C-Phycocyanin From Chaohu Algae

As an important phycobiliprotein with antioxidant properties, cancer-fighting properties and other characteristics, Phycocyanin has widely used as natural pigment, medical and health products and making fluorescent probe in developed countries. In this study, C-Phycocyanin crude extract was obtained by freeze-thaw broken from fresh algae mud in Chaohu, then to explore the efficient extraction and purification technology, the C-Phycocyanin was crude extracted by powdered activated carbon adsorption and purified respectively by aqueous two-phase extraction and salting-out.In the experimental study of C-Phycocyanin crude extraction by powdered activated carbon adsorption, species and particle size of powdered activated carbon adsorption were filtrated firstly; conditions of powdered activated carbon were studied, like temperature, amount, processing time and pH; the optimum conditions of powdered activated carbon adsorption are using coal activated carbon, mesh 400～500, temperature 25℃, amount 100g/L, time 10min and pH 7.0.In aqueous two-phase extraction experiments, the best aqueous two-phase system was selected by analyzing the extraction results of 20 kinds of polymer-salt aqueous two-phase systems, with C-Phycocyanin crude extract treated by powder activated carbon as raw material; then, the optimum conditions of aqueous two-phase purification, which were decided by single factor and orthogonal experiments, are using PEG 4000 and potassium phosphate as phase materials, the concentration of PEG 40004%, the concentration of potassium phosphate 16% and pH 7.0.In the experimental study of C-Phycocyanin purification by multi-step salting-out, effects of ammonium sulfate concentration and salting-out times on purification of C-Phycocyanin were studied by single factor experiments, with C-Phycocyanin crude extract treated by powder activated carbon as raw material; then, with the ammonium sulfate concentration from the first to fourth step as factors, the optimum conditions of four-step salting-out purification were studied by orthogonal experiments. Experimental results show that, the optimum conditions of our-step salting-out purification are 1.0mol/L,2.0 mol/L,1.2mol/L and 2.0mol/L ammonium sulfate concentration. The verified experiment after conditions optimization show that the purity of C-Phycocyanin can reach to the internationally recognized fluorescence reagent grade standard(4.0).