| Agaricus blazei Murill, also known as Agaricus, Komatsu mushroom, is a rare edible and medicinal mushroom. The biological activity of polysaccarides which extract from fermentation broth and mycelium are mainly including antioxidant, anti-aging, anti-tumor, hypoglycemic, hypolipidemic, anti-fatigue and enhance the immunity. Organic germanium has excellent biological functions, including anti-tumor, anti-aging, hematopoietic function, hypolipidemic and inprove the body’s immune system.The extraction conditions of A. blazei exopolysaccharide (EPS), intrecellular polysaccharide (IPS) and intracellular germanium polysaccharide (IGPS) in submerged culture were optimized on the basis of Plackett-Burman (PB) design and response surface methodology (RSM) in this paper. The anti-aging experiments in vivo were done with these three crude polysaccharides. Polysaccharide fractions, obtained after separation and purification of crude polysaccharides, were used to determine antioxidant activity in vitro, monosaccharide composition and chemical structure.(1) The optimal conditions of three A. blazei polysaccharides extractionThe three most significant factors influence on the amount of A. blazei polysaccharide extraction were determined by PB experiments. RSM result shows that extraction of the theoretical amount of EPS, IPS and IGPS were1145.87mg/L,7.52%and7.16%, in their optimum extraction conditions respectively. The results showed that the actual yields of EPS, IPS and IGPS were1127.23mg/L,7.46%and7.12%respectively.(2) Isolation and purification of A blazei polysaccharide fractionsAfter DEAE-cellulose ion exchange separation column and dextran G-100gel column, EPS obtain component EPS-1-1, EPS-1-2and EPS-2, IPS obtain component IPS-1and IPS-2, IGPS obtain component IGPS-1, IGPS-2-1, IGPS-2-2, IGPS-3-1and IGPS-3-2, respectively.(3) The in vitro antioxidant of A. blazei polysaccharide fractions The in vitro inhibition effects of EPS-2at a dosage of200mg/L on DPPH and hydroxyl radicals were32.96±1.65%and96.26±4.81%, respectively. The reducing power of EPS-2was0.98±0.05. The in vitro results showed that EPS-2had very strongly reducing power and scavenging effects on DPPH and hydroxyl radicals.The in vitro inhibition effects of IPS-2at a dosage of1000mg/L on DPPH and hydroxyl radicals were35.94±1.80%and41.2±2.06%, respectively. The reducing power of IPS-2was0.53±0.03. The in vitro results showed that IPS-2had strong reducing power and scavenging effects on DPPH and hydroxyl radicals.The in vitro inhibition effects of IGPS-3-1at a dosage of1000mg/L on DPPH and hydroxyl radicals were46.07±2.30%and61.16±3.06%, respectively. The reducing power of IGPS-3-1was0.60±0.03. The in vitro results showed that IGPS-3-1had strong reducing power and scavenging effects on DPPH and hydroxyl radicals.(4) Anti-aging active of A. blazei polysaccharides in vivoMice were injected aging agent and gavaged A. blazei polysaccharides everyday. The results suggested that EPS, IPS and IGPS could significantly improve the levels of T-AOC in liver tissue of mice, decreased the conteng of MDA in liver tissue and ALT in blood. Compared with the model group, the results of the test group polysaccharide reached a significant level, indicating EPS, IPS and IGPS have a strong anti-aging activity.(5) Monosaccharide composition and molar ratio of A. blazei polysaccharidePolysaccharide fractions (EPS-2, IPS-2and IGPS-3-1) were determined their monosaccharides composition and molar ratio by GC. The result showed that EPS-2was constituted with rhamnose, mannose and glucose, and their molar ratio was1.3:1:2; IPS-2was constituted with rhamnose and mannose, and their molar ratio was1.4:1; IGPS-3-1was constituted with rhamnose, mannose and glucose, and their molar ratio was0.75:0.7:1.(6) Chemical structure determination of A. blazei polysaccharide fractionsPolysaccharide fractions (EPS-2, IPS-2and IGPS-3-1) were scaned by IR in the4000-400cm-1section, the result showed that they were D-pyranoid. |
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