Preparation And Bioactivity Of Protein-bound Polysaccharide From Cultural Medium Of Ganoderma Sinensis

Cancer is a serious threat to human life, it's a common and frequently-occurring disease. More than 10 million's new cases and more than 6 million's deaths occur each year in the world. The mortality rate is in second column, while cardiovascular disease is still stand in the top place.The present study has confirmed that Ganoderma polysaccharides Peptides have significant anti-angiogenesis activity of tumor cell growth also has a certain inhibitory effect, and relative to the polysaccharide, with oral effectiveness of the anti-tumor has a very bright future prospects. Ganoderma is endemic to China; few studies have reported it at home and abroad.This searching project further improves the studies of Ganoderma lucidum antitumor function on the basis of the previous studies. The deep fermentation prescription liquor for Ganoderma sinensis bacterial strains is optimized, and by one-factor-at-a-time methodology and orthogonal experiment, the optical deep liquid fermentation culture medium is determined (w/v) : glucose 3.5%, peptone 0.3%, yeast powder 0.1%, KH2PO4 0.1%, MgSO4 0.05%, vitamin B1 0.005%; The optimal fermentation cultivation condition is: 28°C, 160rpm/min, medium volume 150mL, strains grows best after 14 days, then hypha mycelium yield can reach 6.7 g/L. Removing the floating protein out of the fermentation liquor supernatant by two ways respectively. Different concentrations of TCA and (NH4)2SO4 salting-out results show that protein washout rate and polysaccharide loss rate are both positive relative with TCA and (NH4)2SO4。The protein elution rate will up to 70%, when the concentration of TCA reaches 10%. The results of SDS-PAGE are proving it indirectly. But, meanwhile the polysaccharide percentage loss will increase with protein elution rate growth. It appears a distinct red line on PAS-positive, showing that loss a lot of polysaccharide, over 25%. However, the (NH₄)₂SO₄ (the concentration is 90%) could wash most of protein and control the loss of polysaccharide under 20%. It benefits for protein isolation process. The fermentation supernatant fluid by salting out use the concentration is 90% (NH₄)₂SO₄,Throught the desalination, precede DEAE-Sepharose Fast Flow anion exchange chromatography, get the single sugar peak and single protein peak. In this case, the height of the suger peak and protein peak are superposition. Then, collect the coincidence peak, accquire the purified exocellular protein-bound polysaccharide (EP1). This kind of complex has a obvious Anti-free radical activity and anti-tumour functions.