| 9 evaluated enzymes were selected to extract polysaccharide from fungal mycelia, and then uniform design was utilized for optimaization of technology for high yield polysaccharides from Hypsizigus marmoreus mycelia. the anti-tumor and antioxidant activity of polysaccharide from fungal mycelia was discussed, and the bio-activity of the polysaccharide intro-mycelia and in the wall of mycelia was also discussed.It can get the hignest yield polysaccharides from Hypsizigus marmoreus, Suillus sp and Mycoleptodonoides.sp mycelia by lywallzyme, 14.78±0.22 mg/g, 16.66±1.60 mg/g, 27.49±1.89 mg/g, 164.36%, 156.68%, 141.19% higher than control. It was concluded that Neutral protease A was the most cost efficient among 9 evaluated enzymes when extraction of polysaccharide from mycelia of Hypsizigus marmoreus and Suillus sp, 8.52±0.27 mg/g, 20.67±1.86 mg/g, 56.48%, 44.85% higher than control.The single factor and uniform design experiment was conducted to optimize the conditions, and a regression model were constructed to predict polysaccharide yield from Hypsizigus marmoreus mycelia. The highest polysaccharide yield was obtained when 2% (w/w) of neutral protease was added into the hydrolytic reaction and incubated for 4 h at 43oC, and then extracted one time at 100 oC for 1 h. Using the regression model, the predicted polysaccharide yield was 16.65 (mg/g DMW), which was very close to the measured value of 15.73±0.15 (mg/g DMW). Thus the prediction equation was validated to accurately estimate polysaccharide yield. When following the optimized procedures, polysaccharide yield was raised 75.0% from that prior to optimization. Compared to hot-water extraction and microwave assisted extraction methods, enzymatic extraction method increased polysaccharide yield by 122.5% and 104.8%, respectively. It was concluded that enzymatic extraction is a simple, high yield method to prepare polysaccharide from Hypsizigus marmoreus mycelia.The anti-tumor activity of polysaccharides from Hypsizigus marmoreus mycelia by different enzymatic hydrolysis.has been observed. The test indicated that the anti-tumor activity of polysaccharides extracted by different enzymatic is different. The anti-tumor activity of polysaccharides from Hypsizigus marmoreu mycelia by pectinase, cellulase and acid protease is higher than by other enzymes. The polysaccharides from mycelia by pectinase, cellulose, acid protease and neutral protease 1 can obviously kill the tumor cell of SGC7901, 0.343 mg/mL, 0.397 mg/mL, 0.430 mg/mL, and 0.984 mg/mL respectively in 50% death rate. The anti-tumor activity of polysaccharides in mycelia is higher than polysaccharides from the wall of mycelia.The anti-tumor activity of polysaccharides from Mycoleptodonoides sp mycelia by cellulose B is higher than by other enzymes, it can obviously kill the tumor cell of SGC 7901 in 50% death rate with the dosage of 0.863 mg/mL. The antioxidant activity of polysaccharides from mycelia by different enzymatic hydrolysis.is also different. The antioxidant activity of polysaccharides from mycelia by pectinase, papain, neutral protease A and neutral protease B is higher than by other enzymes. Polysaccharides extracted by papain pectinase, neutral protease A and neutral protease B can obviously scavenge hydroxyl free radical, 51.98μg/mL,55.76μg/mL , 62.49μg/mL , 63.56μg/mL respectively in 50% scavenging rate. Polysaccharides extracted by them can obviously scavenge superoxide anion free radical, 153.77μg/mL,195.61μg/mL,219.85μg/mL,252.68μg/mL respectively in 50% scavenging rate.The antioxidant activity of polysaccharides in mycelia is higher than polysaccharides from the wall of mycelia. The antioxidant activity of polysaccharides from Mycoleptodonoides sp mycelia by papain is higher than by other enzymes, it can obviously scavenge hydroxyl free radical and superoxide anion free radical, with the dosage of 59.42μg/mL, 74.78μg/mL respectively in 50% scavenging rate.
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