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Separation, Purification And Structure On Polysaccharides From Coriolus Versicolor By Fermentation

Posted on:2013-09-14Degree:MasterType:Thesis
Country:ChinaCandidate:J SunFull Text:PDF
GTID:2231330374973095Subject:Biochemical Engineering
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Coriolus versicolor is word wide distributed medicinal fungi, which has become hotspot because of its wide range of pharmacological activity. Polysaccharides produced by versicolor due to its significant hepatoprotective and anti-tumor medicinal activity, are often as clinical assistant drugs. Yields, structures and activity of polysaccharide varied in different media and condition. Therefore, this study is on the screening of the culture medium, cultivating condition, the difference of separation and purification process, and analysis the structure of polysaccharide. The main results are as follows:(1) The effects of various carbon and nitrogen sources. It screens strain YK-505composite carbon and nitrogen sources and the optimization medium by orthogonal test. The optimal fermentation conditions were obtained:Medium:2%glucose,1%sucrose, yeast extract0.15%, peptone0.25%,0.15%potassium dihydrogen phosphate, dipotassium hydrogen phosphate2%,0.15%magnesium sulfate heptahydrate.(2) Compared to submerge fermentation, the solid medium was showed to yield a very lower extracellular and intracellular polysaccharide. Compared to static liquid culture, the submerge fermentation has the ability to shorten fermentation time and improve the yield.(3) Single factor of fermention conditions for biomass production were optimized. The results showed optimal culture conditions were as follows:initial pH, temperature, fermentation time, broth content and rotation speed were6.0,28℃,10.0d,50ml,150r/min, respectively. Response surface methodology was employed for the further optimization. The optium composition conditions of medium time10.0d, broth content60ml and temperture28℃provided the highest biomass production of9.718mg/ml.(4) Single factor of fermention conditions for extracellular polysaccharide production were optimized. The optimal culture conditions were as follows:initial pH, temperature, fermentation time, broth content and rotation speed were5.0,28℃,2.5d,50ml,180r/min, respectively. Response surface methodology was employed for the further optimization. The optium composition conditions of medium time2.5d, broth content68ml and initial pH5.0provided the highest extracellular polysaccharide of12.14mg/ml.(5) Intracellular polysaccharide of high purity (by PAGE) and yield (above24%) are obtained by taking orthogonal design of extraction process, choosing reasonable process condition and further purifying on the base of tradition extraction process. Homogeneous intracellular polysaccharide YK-1is obtained by futher purity and isolation which has high account for56.12%. YK-1mainly is composed of rhamnose, fucose, galactose, glucose, mannose and it contains β-1,3/(3-1,2glycosidic linkage. Extracellular polysaccharide is obtained after the separation, purification, and it has non-peptides, contains β-1,3glycosidic linkage. The monosaccharide composition of extracellular polysaccharide contains at least glucose and xylose. Changes in the absorption maximum of the congo red-polysaccharide complex at various concentrations of hydroxide solution show that the extracellular polysaccharides and YK-1have triple helix form of structure.(6) Intracellular polysaccharide and ginkgo flavone compound can improve the CCl4caused about acute liver injury.There are synergies and dose-effect relationship, can reduce the liver index. Intracellular polysaccharide200mg/kg/d complex with ginkgo flavonoids100mg/kg/d, reduced TBil content, and significantly reduced the ALT and AST activity. ALT and AST were decreased by36.13%,28.89%.
Keywords/Search Tags:Coriolus versicolor polysaccharide, preparation processes, separation andpurification, structure analysis, pharmacological activity
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