Study Of Bioactive Substances Of Fermentation By Pholiota Squarrosa AS5.245 Strain

The edible and medicinal fungi have been used as pharmaceutical pruducts in the world. It is well known that the biologically active substances derived from the fungal species, especially polysaccharides, are regarded as biological response modifier to stimulate human immune function. At present, the edible and medicinal fungi comprise an important source for modern medicine to pick out the powerful new medicine with antitumor and immunostimulating properties. The three-level three-factor Box-Behnken design was executed to optimize the fermentation of AS5.245 and the processes precipitating products of it.The main results of this paper are as follows:1. The three-level three-factor Box-Behnken design was executed to optimize the temperature, time and volume of inoculum during the process of fermentation of AS5.245. And the result is that the volume of inoculum, time and temperature of fermentation of AS5.245 were 12.57mL, 7.82d and 28.57℃ . Respectively, the yield of extra-polysaccharide could reach over 853.727ug/mL.2. Through an orthogonal design, the highest yield was achieved when the sample was applied and disposed for 40 minutes with ultrasonic and for 3 hours in boiling, and the final content of alcohol is 70%.3. The Box-Behnken design was used to the optimization of situation for extraction and separation of extra-polysaccharides produced by pholiota squarrosa, and results showed that the optimal condition to separation was temperature 13.94℃, time19.99h and the final content of alcohol 84.71%. Under the best extraction condition, the highest purity of the polysaccharide could be obtained at 11.56%.4. The crude extra- polysaccharide by AS5.245 was further purified with column chromatography on DEAE Sephadex A-50 fast flow. And the products were identified by the column chromatography on Sephadex G-100 and UV in the range of 190-400nm; the polysaccharide isolated was identified as homogeneous one. Furthermore, the relative molecular weight (MW) is 32,597. And the extra-polysaccharide of AS5.245 is composed of Mannose, Glucose and Galactose with the molar ratio of 3:2:1 in components identified by GC.5. The immunomodulation activities of inter and extra polysaccharides and crude protein from fermentation of AS5.245 were determined. The results showed that the thymus index of three components were from 106.26 ± 15.46 to 129.21±25.99, from 127.92 ±...