A Comparative Study Of Composition And Polysaccharide In Fruiting Bodies And Submergedly Cultured Mycelia Of Grifola Frondosa

Grifola frondosa is a rare medicinal and edible fungi, and with important food andmedicinal values. The cultivation industry of Grifola frondosa developed rapidly withpeople’s needs increasing. In the other hand, the process and product quality of Grifolafrondosa submerged fermentation can been controled easily, and do not need to occupy theland, consumption of forest, and short production cycle advantage during product, so it hasbeen also developed rapidly. Currently, the study of Grifola frondosa nutritional andpharmacological mainly in the fruiting bodies, less about mycelium, but the research ofmycelia nutrition and pharmacologically active substances is very important, and is closelyrelated to the reasonable application of Grifola frondosa fruiting bodies and fermentedmycelia.In this paper, one Grifola frondosa strain with excellent performance of fermentation wasscreened after the molecular identification of4“Grifola frondosa” strains. Then the strainwere cultivated and fermentated with50L fermenter, to prepare fruiting bodies andfermentation mycelia, and the non-volatile components, the volatile components andpolysaccharides of fruiting bodies and mycelia were studied, the main conclusions are asfollows:1.4"Grifola frondosa" strains were determined with molecular identification method,the results showed that3strains were Grifola frondosa, one is Ganoderma lucidumone. oneGrifola frondosa strain with excellent performance of fermentation was screened after themolecular identification of3identified Grifola frondosa strains;2. The mycelia and fermentation broth lyophilized powder yield11.93g/L and28.9g/Lafter fermentation with50L fermenter, respectively. The formation process of fungus ball andthe shape variation of fungus block and fungus ball was observed and found thatmorphological changes can be important supplementary indicators as indicators of pH,biomass, dissolved oxygen during fermentation process;3. The cultivation effects of six formulations of cultivated material were compared in theprocess of laboratory cultivation of Grifola frondosa, the results show that the broad-leavedwood-based formulation was more appropriate to Grifola frondosa growth and the rate ofbiotransformation was more than40%, while those cultivated material added cassava residue,monosodium glutamate residue, or other industrial waste material were not suitable for thegrowth of Grifola frondosa;4. The shape of liquid, solid cultured mycelium and fruiting body mycelium wereobserved with scanning electron microscope. There were significant differences in thickness,homogeneity between three kinds of mycelium. The liquid cultured mycelium containdiaphragm, dose not contain lock-like joint; while the mycelium in fruiting bodies contain not only diaphragm but also lock-shaped joint, and has a typical basidiomycete spore structure;5. We have studied the composition of Grifola frondosa fruiting bodies and mycelia andfound that: there are significant differences in the composition of the non-volatile componentsbetween Grifola frondosa fruiting bodies and mycelia. The content of crude protein, crude fat,reducing sugar, total sugar, free amino acids and hydrolyzed amino acids in mycelia washigher than those in fruiting bodies, while the content of crude fiber, unsaturated fatty acidand flavor nucleotide content was higher in fruiting bodies.62kinds of volatile compoundswere identified from the fruiting bodies, mainly alcohols, ketones, aldehydes; and94kinds ofvolatile compounds were identified from mycelia, mainly aldehydes, ketones, alcohols,esters;6. The refined polysaccharides of Grifola frondosa fruiting bodies and mycelium werepreparated with in addition to fat, boiling water extraction, removal of protein, H₂O₂bleaching, dialysis, freeze-drying, and the yield rate was1.30%and1.40%, respectly. Theyield rate of refined polysaccharide preparated from the crude polysaccharide of ultrafiltratedfermentation broth. The suitable ratio of solid to liquid is1:30and the optimal number ofextraction is3times during the extraction of Grifola frondosa fruiting bodies polysaccharide;7. On DEAE Sepharose Fast Flow and Sephadex G-200column chromatography,homogeneous polysaccharides. GF04FIPS01, GF04FIPS02were obtained from fruitingbodies; GF04MIPS01, GF04MIPS02, GF04MIPS03, GF04MIPS04, GF04MIPS05,GF04MIPS06, GF04MIPS07from mycelia; GF04EPS01, GF04EPS02from the fermentationbroth, and determined the molecular weight, polysaccharide content, protein content, uronicacid content, monosaccharide composition, the results showed that: the reconstitution capacityof10homogeneous polysaccharides are different, GF04MIPS05, GF04MIPS06, GF04FIPS02are better; GF04EPS02, GF04MIPS02are poor, the protein content of10samples are between0.84%-5.74%and uronic acid are4.41%-21.90%. The molecular weight of GF04FIPS01,GF04FIPS02are6.03×10⁵,4.70×10⁵, and contain glucose, galactose, fucose, mannose; themolecular weight of GF04MIPS01, GF04MIPS02, GF04MIPS04, GF04MIPS05,GF04MIPS06, GF04MIPS07are from2.03×10⁴to1.76×10⁶, GF04MIPS01, GF04MIPS01containing glucose and a small amount of mannose, the other component only containsglucose; the molecular weight of GF04EPS01, GF04EPS02are1.50×10⁴,1.91×10⁴,GF04EPS01contains glucose, fucose, mannose, galactose, and GF04EPS01contains glucose,fucose, mannose, galactose, arabinose, xylose;8. Infrared studies show that all10kinds of homogeneous polysaccharides are pyranosepolysaccharides, GF04FIPS02, GF04MIPS04GF04MIPS05, GF04MIPS07GF04EPS01onlycontain α-glycosidic bond, GF04FIPS01, GF04MIPS01, GF04MIPS02, GF04MIPS06,GF04EPS02containing bothα-,β-glycosidic bonds. After1H-NMR and¹³C-NMR analysis,we obtained that GF04FIPS02contains→3)-α-D-Manp-(1→, α-D-Galp-（1→, →6）-α-D-Glcp-(1→, α-L-fucp-(1→; GF04MIPS01contains β-D-Glcp-（1→,→3）-β-D-Glcp-（1→,→3）-α-D-manp-（1→; GF04MIPS05contains→3）-α-D-Glcp-（1→,→3,6）-α-D-Manp-（1→; GF04EPS01contains→3）-α-D-Manp-（1→,→3）-α-D-Glcp-（1→,→3）-α-L-Fucp-(1→.