Fermentation And Immunoreactivity Of Extracellular Polysaccharide By P.baumii Pilát

Medicinal fungus has been used as a traditional Chinese medicine in China for several thousand years because of their various biological and pharmacological active ingredients, especially, polysaccharides. Recently, polysaccharides from medicinal fungus have been well-known as a potential Biological response modifier. Modern pharmacological studies showed that medicinal fungi polysaccharides have immunostimulating and anti-tumor properties."Sanghuang", which has been used as a traditional Chinese medicine in China for a long time, has gained wide popularity as potential immunostimulating and anti-tumor properties and has become one of the valuable medicinal fungi in the world. Therefore, it deserves investigation to produce EPS from P. baumii Pilat by submerged culture and immunostimulating activities of EPS. The main results are shown as follows:1. A 2(7-3) fractional factorial design (FFD) was employed to determine the key ingredients significantly affecting EPS production from P. baumii Pilat. Seven nutrient factors considered for the design were glucose, peptone, yeast extract, diammonium oxalate monohydrate, KH2PO4, MgSO4 and thiamine (VB1). Among the variables screened, the three variables including glucose, yeast extract and diammonium oxalate monohydrate were determined as the most significant variables influencing EPS production.2. The levels of the significant parameters and interaction effects between various medium components which influence significantly the EPS production from FFD were analyzed and optimized by using a central composite design (CCD) in response surface methodology (RSM) and the response surface regression procedure. The optimal medium composition was determined as following (g/1):glucose 34.12, yeast extract 5.01, peptone 4, MgSO4 0.75, KH2PO4 1, VB1 0.0075 and diammonium oxalate monohydrate 0.88.3. The effect of EPS from P. baumii Pilat on the splenocyte proliferative response in vitro was investigated by using MTT-based colorimetric method. In the absence of mitogen, EPS at 100μg/ml enhanced the proliferation with the highest splenocyte proliferation index (1.18±0.09). Meantime, the results also showed that EPS could enhance both the LPS-induced B cells and Con A-induced T cells proliferation.4. The ability of EPS from P. baumii Pilat to stimulate acid phosphatase activity in peritoneal macrophages in vitro was investigated by using the cell model experiments. Results showed that the activities of acid phosphatase in macrophages increased significantly by EPS in a dose-dependent manner (25-200μg/ml). Furthermore. EPS at 200μg/ml activated the acid phosphatase activity by 105.50% of the control. These results suggested that the EPS from P. baumii Pilat strongly induced the activation of macrophage, increased significantly acid phosphatase activity in peritoneal macrophages. and possessed higher immunomodulatory potential.