Optimized extraction and purification technology of Gekko sulfated glycopeptide based on its anticancer activities

Objective:To optimize the extraction and purification technologies of Gekko sulfated glycopeptide based on the content of glycopeptide, removal ratio of proteins, and anticancer activities. Methods: Different extraction methods, namely, water extraction, ultrasonic extraction, enzymatic hydrolysiswater extraction, and enzymatic hydrolysisultrasonic extraction were considered to determine the best extraction method. Single factor and orthogonal experiments were performed to determine the optimum extracting conditions. Sevage, enzymatic hydrolysis-Sevage, trichloroacetic acid (TCA), TCA-Sevage and enzymatic hydrolysis-TCA methods were tested to determine the best deproteinization method. The glycopeptide content and protein removal ratio were analyzed by the phenolsulfuric acid and Coomassie Brilliant Blue methods. Results:Gekko sulfated glycopeptide obtained by water extraction could inhibit the proliferation of HepG2 and SKBR3, as well as promote that of lymphocytes. The glycopeptide content was 4.049% in the optimal extracting condition of a triple decoction extraction for 1 hour each time with a material to solvent ratio of 1:15. The enzymatic hydrolysis-TCA method was found to be the optimal deproteinization method, with a protein removal ratio of 50.46%, glycopeptide content of 14.27%, and inhibitory ratio on human HepG2 cells of 49.06%. Conclusion:This extraction and purification technique for Gekko sulfated glycopeptide is reasonable, feasible, and provides a scientific basis for industrial production.