| Ganoderma lucidum is a valuable edible-medicinal fungus in our country.The polysaccharide,triterpenoids,flavone are the important active ingredient of Ganoderma lucidum,whose biological activity attracted more and more attention.In order to explore the potential value of functional food materials and drug leads,different extraction methods of polysaccharide,triterpene,flavone in Ganoderma lucidum werestudied.And then isolation and purification were carried on to obtain pure active compound.Finally,the structure of pure polysaccharide,triterpene,flavone and the activity on antioxidant were discussesed.The main results are as follows.1.Through comparative analysis of water extraction,alcohol extraction and water extraction after alcohol extraction,it showed that alcohol extraction after water extraction methods would be better for the extraction rate of the active compounds in Ganoderma lucidum.And orthogonal test showed that:optimal extraction conditions of polysaccharides,extraction time 2.5h,extraction temperature 95℃,solid-liquid ratio 1:25,the polysaccharide yields 2.09 mg/g;optimum extraction conditions of triterpenoids:65%ethanol,extraction temperature 75 ℃,extraction time 1.5h,solid-liquid ratio 1:20,triterpenoids yields were 1.019mg/g;optimal extraction conditions of flavonoids:alcohol extraction,65%ethanol,extraction temperature 75℃,extraction time 1.5h,solid-liquid ratio 1:20 flavonoids yields were 0.56 mg/g.2.On the basis of the above results,extraction rate of the active compounds in Ganoderma lucidum by means of multiple enzymes wrre optimized using the software Design Expert.Response surface experiments showed that:the optimal enzyme assisted extraction conditions of Ganoderma lucidum polysaccharides:cellulase addition 1.55%,pectinase addition 1.06%,neutral protease addition 2.55%,polysaccharides the extraction rate up to 2.47 mg/g;optimal enzyme assisted extraction conditions of triterpenoids:cellulase addition 2.07%,pectinase addition 0.87%,neutral protease addition 2.06%,triterpenoids yields were 1.319mg/g;optimal enzyme assisted extraction conditions of flavonoids:cellulose enzyme addition 1.52%,pectinase addition amount 1.21%,neutral protease addition 2.48%,flavonoids were 0.75 mg/g.3.The polysaccharide was purified by DEAE Sephadex A-50,four pure polysaccharide components,named GLPS-1、CLPS-2、GLPS-4、GLPS-5,were obtained.Three triterpenoids components were purified by silica gel column chromatography,,named TP-1、TP-2、TP-3、TP-4.Three flavonoid components were gained by silica gel column chromatography,,named FP-1,FP-2.4.Antioxidant activies of the puried compounds:Results of the polysaccharids on DPPH clearance rate showed that enzyme extract methods has the highest antioxidant activity,when the amounts of polysaccharides reached 3.2mg/ml,the DPPH clearance rate reached 85.2%;purified polysaccharide components exhibited a certain amount of scavenging,when the polysaccharide is 2.6mg/ml,GLPS-5 showed the highest DPPH clearance rate,about 50.4%;Enzyme extraction of Ganoderma lucidum polysaccharides also showed strong reducing power,when 10mg/ml polysaccharides force reaches 1.928.Of which,the reducing power of GLPS-5 was about 1.021.Results of the triterpenoids on DPPH clearance rate showed that alcohol extraction was the optimal method for antioxidant triterpenoids,when the extraction amount was2.4mg/mL,the DPPH clearance rate reached 77.6%.Some purified components showed certain DPPH clearance rate,the DPPH clearance rate of TP-1 is the highest(38.8%)when the amount of triterpenoids reached 2.4mg/ml.Hydroxyl radical clearance rate of Ganoderma triterpenoids showed that alcohol extraction has the highest antioxidant active,when extracts of triterpenoids is 2.4mg/ml,the triterpenoids clearance is 61.4%,Some the purified components have shown a certain amount of scavenging,when triterpenoids was 2.4mg/ml,TP-1 clearance rate was higher,which is 27.7%;alcohol extract of Ganoderma triterpenoids showed strong reducing power,when reducing power in 9mg/ml the active reached 1.838,and the purified components TP-1,TP-2,TP-3,TP-4 reducing power reached 1.01,0.911,0.743,0.661.Ganoderma flavonoids DPPH clearance rate results,alcohol extract antioxidant flavonoids is the highest,when the flavonoids is 2.6mg/ml,the clearance rate reached 57.1%.Purified components showed some scavenging,when flavone is 2.4mg/ml,FP-1 clearance is higher,which is 32.2%.Ganoderma flavonoids on hydroxyl radical scavenging results,alcohol extraction flavonoids has the highest antioxidant activite.when the extract flavonoids is 2.6mg/ml,crude flavonoids clearance rate reached 61.1%.The purified components showed some scavenging,when the flavone is 2.4mg/ml,FP-1 clearance is higher than others,which is 34.2%.Alcohol extraction flavonoids showed strong reducing power,when the amount is 9mg/ml,it reached 1.738.The purified compounds FP-1 and FP-2 reducing power are 0.673 and 0.611. |
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