Comparison Of Powdery Activated Carbon Adsorption Combined With Column Chromatography And Ultrafiltration For Purifying Phycocyanin

In recent years,the extraction of proteins,amino acids and pigments from cyanobacteria has become a hot spot in the research of biology,food engineering,and life health science.The phycocyanin extracted from the cyanobacteria is widely used.It can be used as fluorescent molecular probe,and it can also be used as nutrient component or pigment for foods and cosmetics.In this study,fresh cyanobacterial mud from Chaohu Lake was used as raw material,and the algae was broke down by freezing and thawing to obtain crude phycocyanin.Then phycocyanin was extracted by powdered activated carbon adsorption,and then phycocyanin was purified by hydroxyapatite column chromatography and ultrafiltration,respectively.The study was to optimize the process combination of extraction and purification of high-purity phycocyanin.In the study of powdered activated carbon adsorption,the type and particle size of the powdered activated carbon for the purification of phycocyanin were screened.Then the particle size,addition amount,phycocyanin concentration,and time were studied.Consequently,the optimum conditions for powdered activated carbon adsorption were using coal powdered activated carbon,mesh 400 ~ 500,phycocyanin concentration 1.0 mg/ m L,addition amount 100 g/ L and time 15 min.In the study of ultrafiltration,after phycocyanin was purified by powdered activated carbon adsorption,the effect of different molecular weight cut-off on phycocyanin purification was studied.Then PC concentration,time,and centrifugal rotation speed were studied.Consequently,the optimum conditions for ultrafiltration were molecular weight cut-off 10 KDa,phycocyanin concentration 0.45 mg/ m L,centrifugal speed 4 000 g,and time 30 min.The purity of phycocyanin could reach the pharmaceutical grade(A620/A280>3.0).In the study of column chromatography,after phycocyanin was purified by powdered activated carbon adsorption,the phycocyanin was purified by hydroxyapatite column chromatography.The p H and concentration of eluent were selected as factors of the single factor experiments.Consequently,the optimum conditions for hydroxyapatite column chromatography were p H 7.0,concentration 25 mmol/L,the purity of phycocyanin could reach the reagent grade(A620/A280≥4.0).