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摘要：灵芝Ganoderma lucidum (Leyss. ex Fr.)Karst在我国是著名的药用真菌，现代研究结果表明灵芝的主要活性成分是三萜类化合物。本研究通过多批次的两阶段培养发酵，制备了足够生物量的灵芝菌丝体。通过HPLC分析了灵芝两阶段发酵获得的菌丝体中的核苷类化合物。结果发现，灵芝菌丝体中含有尿嘧啶；胞苷；次黄嘌呤；尿苷；腺嘌呤；肌苷；鸟苷；腺苷等核苷类化合物。
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将上层菌丝体和下层菌丝体合并，用95%的乙醇溶液提取3次，合并提取液后浓缩。用石油醚，氯仿，乙酸乙酯，正丁醇萃取浓缩物，得到不同组分。通过HPLC分析发现，氯仿萃取部分中含有大多数的灵芝三萜化合物，因此本研究对氯仿部分进行了分离纯化。24235
对氯仿组分F6(24-32)分离得到化合物Ⅰ和Ⅱ。运用现代光谱技术(ESI-MS、1D NMR、2D NMR)对两种化合物的结构进行了鉴定，两个化合物均为已知的化合物。化合物Ⅰ为星鱼甾醇(Ergosta -7, 22- dien-3β-ol)；化合物Ⅱ为灵芝酸S(Ganoderic acid S)。本研究为今后灵芝菌丝体中三萜化合物的活性研究提供了基础。
毕业论文关键词：两阶段培养发酵，灵芝菌丝体，三萜化合物，HPLC
The research in two-stage submerged fermentation of Ganoderma
Abstract：Ganoderma lucidum (Leyss. ex Fr.) Karst. is a well-known medicinal mushroom in China. Triterpenoids are important bioactive constituents of G.lucidum. In this study, the biomass of Ganoderma lucidum mycelium was prepared enough by many batches two-phase culture fermentation.HPLC has analyzed nucleoside compounds from two phase Ganoderma lucidum fermentation mycelium. The results show that the Ganoderma lucidum mycelium contains nucleoside compounds:Uracil、cytidine、hypoxanthine、uridine、adenine、 inosine guanosine、 adenosine.
Incorporated the upper mycelium and the under mycelium, extractied three time with 95% ethanol, incorporated the extracted liquid and concentrated. Extracted the ethanol extraction with petroleum ether, chloroform, ethyl acetate and n-butanol, got petroleum ether, chloroform, ethyl acetate and n-butyl alcohol part . By HPLC analysis,it was found that most of the Ganoderma lucidum triterpenoid compounds consist in the chloroform extraction part . So, in this study ,we separated and purificated the chloroform part.
CompoundⅠand compound Ⅱ were found in F6 (24-32) components isolated from chloroform part for further purification and got, Used the modern spectrum technology (ESI - MS NMR, 1 d and 2 d NMR) to identify the structure of two compounds, they were all known. Compound Ⅰ for Ergosta-7,22- dien -3β- ol; Compound Ⅱ for Ganoderic acid (S).This study provides the basis for the active research of triterpenoid compounds from Ganoderma lucidum mycelium in the future .
KEY WORDS: the two-stage culture method, Ganoderma lucidum mycelium
, triterpenoid , HPLC
目录
1 前言 1
1.1 研究背景 1
1.1.2灵芝 1
1.2 研究现状 1
1.2.1 灵芝中的化学成分 1
1.2.2 灵芝三萜类化合物 2
1.2.3 灵芝三萜类化合物特征 2
1.2.4 液体深层发酵技术 4
1.2.5 三萜类化合物的提取方法 5
1.2.6灵芝三萜类化合物的分离纯化 6
1.2.7 灵芝三萜类化合物的药理活性 7
1.3 研究的目的与意义 8
1.4 研究内容 9
2实验方法和实验内容 10
2.1 实验材料 10