Ganoderma tropicum, a Chinese medical mushroom, has been reported with the value of anti-tumor, treatment of coronary heart disease and protecting the lever. G.tropicum belongs to the Ganodermataceae, Aphyllophorales, Basidiomycotina. Recently, few culture conditions have been carried on to enhance the bioactive components. In this work, by both solid and liquid fermentation approaches, the orthogonal experiment was employed to optimize the medium and obtain the most important medium components on the yield of the bioactive components of polysaccharides and triterpenoids. In chapter 2, the G.tropicum mycelium was firstly fermented by plate medium fermentation and liquid shake fermentation for the accumulation of seeds mycelium. Subsequently, aiming at the mycelium weights, amounts of polysaccharides and triterpenoids respectively, a 3-level orthogonal experiment was carried on to gain the best medium composition. The experiment predicted that the most suitable medium for mycelium weights was potato and sucrose, 10 g yeast extract. The yield of polysaccharides in G.tropicum was investigated by phenol sulfuric acid method and then the optimum conditions of extraction were determined as potato and sucrose and 7.5 g yeast extract. Interestingly, it was proved that there were no triterpenoids in the mycelium, and the supernatant maybe showed triterpenoids produced. To gain the most amounts of triterpenoids in the supernatant, potato + sucrose, 7.5 g yeast extract and 10 mM CaCl2 was calculated as the best medium. Subsequently, based on cultured in the growth phase successfully, the variation regulation was studied on purpose for fruiting body weights and bioactive components of G.tropicum. The culture time was finalized that 26 days fermentation were the most suitable time for G.tropicum weights and polysaccharides production. However, the highest amount of triterpenoids in G.tropicum appeared at 31th day cultivation. 澳門大學碩士學位論文 IV Finally, the medium optimization in growth phase was also carried on by a 3-level orthogonal experiment and cultured both in the incubator and in the wild. In the two places, 1/2 cotton seed hulls and 1/2 sawdust, 50 g gypsum powder and 10 g sucrose was both the optimized medium for the fruiting body weight. In determination of polysaccharides, 1/2 cotton seed hulls+1/2 wood as carbon source, sucrose 10 g were the excellent levels. In terms of both methods, carbon source was the most important factor. In the cultivation, gypsum 10 g was a better choice in the wild, and gypsum 2 g was better in the incubator. There were triterpenoids in the fruiting bodies instead of the medium. 1/2 cotton seed hulls and 1/2 sawdust as carbon source, 50 g gypsum powder and 5 g sucrose were the medium which was the fittest for the triterpenoids of the fruiting body. In this study, we evaluated the pharmacological components by combining biological cultivation and HPTLC method. It is surely that it will provide the basis for future medicinal optimization.

熱帶靈芝 Ganoderma tropicum(Jungh.)Bres 是担子菌亚门（Basidiomycotina）、 非褶菌目(Aphyllophorales)、灵芝科（Ganodermataceae）真菌，具有抗腫瘤、保 肝、治療冠心病等功效。近年來關於熱帶靈芝藥理活性物質培養優化的研究非常 少，所以本實驗採用液體發酵和固體栽培的方法，對熱帶靈芝進行優化培養和研 究。 本文第二章首先通過固體平板培養和液體搖瓶發酵的方法對熱帶靈芝菌絲 體進行發酵培養，并對其進行顯微觀察。之後，通過三因素三水平的正交實驗對 培養基的組成進行優化，繼而分別針對菌絲體產量、多糖含量和三萜產量，確定 最優培養基成份。最終通過實驗確定：以菌絲體菌重為優化參數，則碳源為最主 要影響因素，其次為 CaCl2 和氮源。通過苯酚硫酸法對多糖含量進行檢測，進而 正交分析。確定最適合多糖生成的培養基配方為蔗糖和馬鈴薯，7.5 g 酵母粉。 在靈芝三萜的液體發酵培養方面，發酵菌絲體中並無三萜成份但上清液中卻具有 可能是三萜的成份。經過正交實驗，其優水平依然是蔗糖+馬鈴薯，但以 7.5 g 酵母粉為氮源和 CaCl2 10 mM。碳源是最主要影響因素。 其次，在液體搖瓶發酵的基础上，進一步對熱帶靈芝進行固體栽培研究。第 三章首先對固體栽培熱帶靈芝進行動態累積的測定，即採摘不同培養時長的熱帶 靈芝子實體(相同培養條件)并對其多糖三萜成份進行測定。結果顯示，在開蓋培 養第 26 天，熱帶靈芝子實體產量和多糖產量同時達到最大值。在開蓋培養第 31 天，熱帶靈芝子實體中三萜含量達到最高。而固體栽培培養基中無三萜類物質。 第三章通過室內培養箱培養和室外培養的方法，通過三因素三水平的正交實 驗分別對固體栽培培養基進行優化。在室內和室外培養的兩種培養環境中，以子 實體重量為優化參數，最优培養基均为 1/2 棉籽殼+1/2 木屑为碳源、50 g 石膏粉、 10 g 蔗糖，碳源為最主要影響因素。在多糖優化方面，採用苯酚硫酸法對粗多糖 進行含量測定。適合粗多糖累積的優水平分別為碳源：1/2 棉籽殼+1/2 木屑，蔗 糖 10g。对两种培养方式而言，碳源是最主要的影响因素。但是在石膏粉方面， 室內栽培的最優水平為 2 g 而室外的最優水平為 10 g。就三萜而言，固體栽培實 驗子實體中含有三萜，而培養基中無三萜。兩種不同培養分別進行優化，最優三 萜產量組合均為 1/2 棉籽殼+1/2 木屑为碳源、50 g 石膏粉和 5 g 蔗糖。 澳門大學碩士學位論文 II 本研究使用生物培養和 HPTLC 相結合的方法對熱帶靈芝藥理活性成份進行 評估，為科學培養和栽培熱帶靈芝提供依據，為今後中藥成份優化提供了參考。