| Coriolus versicolor is a world wide distributing medicinal fungi which is sweet and cool, can clear away heat and eliminate inflammation, and whose indications of tracheitis, hepatitis and tumor. Its neoteric research of medicinal value is based on folk experience. With unceasingly further study, ceaseless improvement of research level and extension of application range researcher put forward that it has supplement orientation of the effect of supporting the healthy energy , invigorating energy and replenishing qi. In order to develop submerged fermentation technique and to reach high level of industrical production of Coriolus versicolor, the following aspects, including screen good strain and submerged culture process.The conditions of protoplast formation and regeneration from the mycelia of Coriolus versicolor were studied. Experimental results indicated that the highest production rate of protoplast can be obtained by using mycelial age of 3d, 0.4mol/L KCL, 3% lywallzyme digesting for 5 hours at the condition of 30℃and pH 5.5. The highest regeneration rate was obtained on RM1 medium layers.By means of ultraviolet mutation, a mutant strain YBUV113 was screened. The strain was well grown in PDA solid-state medium. It produces almost the same amount of mycelia and PSP(polysaccharopeptide of Coriolus versicolor) as the original strain did when grown in liquid state medium.Carbon and nitrogen sources are important factors that effected the growen of mycelium and yield of polysaccharopeptide of Coriolus versicolor. The optumum carbon source is maize powder. The optumum nitrogen source is soybean oil meal. The suitable shaking culture conditions are as follows: initial pH of medium 5.5, inoculation quantity 20%, shaking frequency 160r/min, incubation time 3 days. Under this condition, the maximum mycelia dry weight is about 34.5g/L. Intracellular polysaccharide of 53.74% and protain of 18.36% were obtained.The Prescription of Ziseng Zhenggan is composed of polysaccharidopeptide, salvia, panax notoginseng, red peony root, kudzu root, ginkgo biloba leaves and so on. This prescription is a self-drafted prescription based on the principle"strengthening body resistance and driving away evil, promoting blood flow and removing stasis"and consulted on some experiences of clinical medication. This prescription is also a modern Chinese traditional medicine compound preparation (including six Chinese traditional medicine drugs) with independent innovation. The study on pharmacodynamics has proved that this prescription can strengthening body resistance, promoting blood flow and remove stasis, decrease serum lipids, prevent fatty liver disease, prevent liver fibrosis, counteract hepatitis B virus and so on. Ziseng Zhenggan can be also applied to treat fatty liver disease and liver fibrosis. We choose the pattern number of macroporous resin By the method of static-state adsorption and desorption experiments, And by different extracting methods to compare every experiment, the best extracting method is confirmed. Research the dynamic adsorption of macroporous resin. By desorption experiment, the maximum adsorptive capacity of the macroporous resin and the best elution concentration can be confirmed. By using orthogonal experiment method, the optimal parameters of extracting technology and purified technology are confirmed. As a result, based on 100g as a standard, the best condition of extracting technology includes concentration of ethanol 70%, dosage of ethanol 1400mL, speed of seeping 1mL/min. The best condition of purified technology includes the flow rate of sample 3Bv/h, the volume of ethanol 3Bv, concentration of sample 200mg/mL, the flow rate of elution 3Bv/h. Active ingredients of polysaccharide-peptide,salvia,panax notginseng,red peony root, kudzu root,ginkgo biloba leaves by optimum extratction is 45%. |
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