Isolation, Purification And Structural Characteristics Of Polysaccharides From Ganoderma Lucidum Spore Powder

Ganoderma lingzhi belongs to the family Ganodermataceae and is one of the traditional medical fungi of China. Studies reported that fruiting bodies, mycelia and spore powder of Ganoderma lingzhi had similar functions like antineoplastic and immunoregulation, which was closely related to the content of polysaccharides conteined in them. Recently, studies about spore powder of Ganoderma lingzhi increased with the substantial raise of production of spore powder, most of which were concentrated on the intercellular polysaccharides, while studies on polysaccharides of cell wall in spore powder of Ganoderma lingzhi were rare. The aim of this paper was to intentively study the extraction process, purification,structural elucidation and biological activity of polysaccharides of cell wall in spore powder of Ganoderma lingzhi, supporting theoretical basises for product development of spore powder of Ganoderma lingzhi in the future. This paper was made up of two parts, and related resultes were showed as follows.1. Comparation of polysaccharides and other chemical compositions extracted from fruiting bodies, mycela and spore powder of Ganoderma lingzhiThe differences of polysaccharides, proteins, amino acids, glucose, sugar alcohols, nucleosides and triterpenes extracted from fruiting bodies, mycelia and spore powder of Ganoderma lingzhi were compared. Monosaccharide compositions of the polysaccharides were analyzed by ion chromatography-integrated pulsed amperometric detector, HPLC spectrums and their changes after treated with?-amylase and endo-1,3-?-glucanase were also compared. The data of HPLC fingerprints of the triterpenes extracted was analyzed by similarity. Results showed that contents of polysaccharides, proteins, amino acids, glucose, sugar alcohols,nucleosides and triterpenes in fruiting bodies, mycelia and spore powder of Ganoderma lingzhi had significantly differences. Polysaccharide content of mycelia was 3.81%, which was the highest one, and those of spore powder and fruiting body were 1.8% and 0.59%, respectively. Monosaccharide composition, molar ratio and structure of the polysaccharides from mycelia, fruiting bodies and spore powder were also different. NO release experiment using RAW 264.7 macrophages revealed that both polysaccharides from mycelia and fruiting bodies showed good activity ofstimulating macrophages to release NO, however the polysaccharide from spore powder showed relatively lower activity. Content of Ganoderma lingzhi triterpenoids was peaked in friut bodies, with wide varieties. However content of Ganoderma lingzhi triterpenoids of mycelia was low with less kinds. Meanwhile, triterpenoids in unbroken spore powder of Ganoderma lingzhi were hardly detected. The study demonstrated that the chemical components of these three materials varied significantly.2. Purification and structural elucidation of polysaccharides in cell wall of spore powder of Ganoderma lingzhiSpore powder of Ganoderma lingzhi was carried out wall-breaking treatments by superfine grinding technology after the remove of water-soluble intercellular polysaccharides inside. Then it was extracted by water extraction with alcohol sedimentation to obtain polysaccharides in cell wall after the remove of lipide material. There polysaccharides were purified by DEAE–Sepharose Fast Flow anion exchange chromatography, and Sephacryl-400 gel filtration chromatography. Finally,S400-1 and S400-2 was abtained, while the yield of the latter is higher than that of the former. After purification, the structure characterion of S400-2 was indicated by IR, methylation, NMR and so on. Results showed that S400-2 was just composed by glucose and contained two configurations of ?-glucan and ?-glucan with varied linking methods. Chemical shift assignment of carbon and hydrogen compounds of S400-2 were made according to the spectral data. Besides, biological and moisturizing tests indicated that polysaccharides in cell wall showed well immune-boosting and moisturizing effects.Orthogonal experiment was taken to get the optimized extraction conditions of polysaccharides in cell wall of spore powder by alkalies. Results showed that optimized conditions were: extracted with 1 mo L/L Na OH at 70 oC for 3h and 1:20 of the ratio of material and solvent, provide theoretic foundation for the study of these polysaccharides.