Production Of Inonotus Obliquus Mycelium By Submerged Fermentation And Analysis On Its Active Ingredients

Inonotus obliquus(Fr.)Pilat has been known for over one hundred years as a medicinal fungi in Russian Federation. It can treat many difficult diseases, such as anti-tumor, anti- inflammatory, anti-oxidant, hypoglycemic, hypocholesterolemic immuno-stimulating activities and so on. Inonotus obliquus D was screened in this paper, and the optimum fermentation condition of Inonotus obliquus D for high yield of intracellular and extracellular polysaccharopeptide. The extractive technique of intracellular polysaccharide and triterpenoids from Inonotus obliquus were also studied. The characterization of intracellular and extracellular polysaccharopeptide was primarily identified. The results were as follows:Inonotus obliquus D with high grow speed, dense hyphae and high yield of polysaccharide was screened. Submerged fermentation of Inonotus obliquus D was studied. The optimum fermentation medium were as follows: glucose 2.5%, yeast extract 0.4%, corn meal 0.8%, soybean flour 1.0%, wheat bran 0.7%, K2HPO4 0.1% and MgSO4 0.05%, pH6.0. The optimum fermentation condition were as follows: inoculation amount 10%(V/V), medium load amount 100 mL/250 mL shaking flask, temperature 28℃, culture time 10 days, rotary speeds of shaking flask is 160 r/min. Under the optimum fermentation medium and conditions, Inonotus obliquus D holded rough pellets, and the yield of mycelium dry weight, the yield of extracellular and intracellular polysaccharide of Inonotus obliquus are repectively were 41.12 g/L, 5.95 g/L, 2.93 g/L.Extraction conditions of polysaccharopeptide were obtained, which were as follows: extraction temperature 80℃, the ration of material to water 1:40, extraction time 2 hours, extract 2 times, the voluminal ratio of ethanol to supernate 3:1 and deposition temperature 4℃. Under the optimum extraction conditions, the extraction rate of polysaccharopeptide was 59.5 mg/g. The extractive technique of triterpenoids from Inonotus obliquus was also studied. The optimum extraction condition of triterpenoid were: 80 mL/g isopropanol and the extraction time 2 h with 80℃, Under the optimum extraction conditions, the extraction rate of triterpenoids was 8.85%.The characterization of intracellular and extracellular polysaccharopeptide were primarily identified . The results showed that the extracellular total sugar content was 59.95%, peptide content was 23.81%, reducing sugar content was 0.26%. The intracellular total sugar content was 62.86%, peptide content was 24.51%, reducing sugar content was 0.31%. The molecular weight of valid peak the intracellular polysaccharopeptide is 762.801 KDa and its content is 78.56% in total, the extracellular polysaccharopeptide is 953.005 KDa and its content is 62.23%.There were seventeen kinds of amino acids in intracellular and extracellular polysaccharopeptide, and essential amino acid accounted for 19.23% and 16.60% of the total amino acids, respectively. In intracellular and extracellular polysaccharopeptide, glutamic acid accounted largest, and asptic acid accounted second largest The polysaccharopeptide was scanned by UV from 190 nm to 400 nm, and the figure showed the characteristic of proteglycan.They also showed characteristic ofα-linkages andβ-linkages by analysis with IR.