Studies Of Fermented Ganoderma Lucidum Polysaccharides Modified By Sulfuric Acid And Their Biological Activity

This paper is about the anti-tumor and antioxidant activity of the Ganoderma lucidumpolysaccharide which was extracted from the submerged Ganoderma lucidum fermentation.The extracellular Ganoderma lucidum polysaccharides was sulfation modified aiming toenhancing the antitumor activities.Sulfamic acid that act as the esterification reagent wasused instead of chlorosulfonic acid. Several parameters were investigated including thesolvents, reaction temperature, microwave-assist, ultrasonic assist, and catalyst, in order to gethigh degree of substitution sulfated polysaccharide.1. The extraction rate of polysaccharide was9.46mg/g dry weight of mycelium with themethod of hot water extraction,while the extraction rate was up to13.02mg/g dry weightof mycelium with microwave assisted. Ultrasonic had little effect on the polysaccharidesextraction.2. Synthetic process was studied by single factor experiment on conventional heating andmicrowave assisted heating.Sulfated extracellular polysaccharide of Ganoderma lucidumsubstitution degree can reach1.34under condition of conventional heating,while the DScan reach up to2.63with microwave assisted.3. Antioxidant and antitumor activity was studied on extracellular and intracellularpolysaccharide of Ganoderma lucidum, and different degrees of substitution of sulfatedpolysaccharide.The intracellular polysaccharide inhibits the growth of tumor cells cyclewas analyzed by flow cytometry.Extracellular and intracellular polysaccharide of Ganoderma lucidum, and differentdegrees of substitution of sulfated polysaccharide (DS1.64and2.67) has a certain abilityof scavenging DPPH free radical and hydroxyl radical scavenging ability strong, whilethe inhibition activity of the tyrosine enzyme and lipid oxidation resistance is weak;MTT results indicated that:Extracellular polysaccharide of Ganoderma lucidum had littlle inhibitory activity onHepG2and Bel-7404. Meanwhile, intracellular polysaccharides of Ganoderma lucidumhas certain inhibition effect on Huh-7, Bel-7404, HepG2,4T1, U-20S, and littleinhibitory effect on L-02, H1299.The highest inhibition of HepG2was93.7%with the DS1.64sulfated extracellularpolysaccharide while the DS2.67sulfated extracellular polysaccharide had the inhibitionof64.7%. Meanwhile, sulfated polysaccharide mixing with intracellular polysaccharideof Ganoderma lucidum had30%inhibition of normal liver cell.FlowCytometry indicated that intracellular polysaccharides of Ganoderma lucidum hadeffecton HepG2cells in G2/M phase in24h, and the proportion of cells in the cell cyclewas increased1times,48h,72h did not change significantly; but by adding drugs thatappeared obvious apoptosis peak in the drug group72h. Intracellular polysaccharides ofGanoderma lucidum on L-02cell cycle has little effect on the phase distribution...