臺灣博碩士論文加值系統

樟芝（Taiwanofungus camphorates）為台灣特有的藥用菇類，活性產物對於人體有提升免疫力與保肝等功能。由於樟芝於野地生長緩慢且取之不易，因此近年來使用固態與液態培養，以品質較穩定且價格合理的方式，供應保健食品市場來符合大眾的需求。本實驗探討以幾丁聚醣與樟屬植物之抽出物添加於固、液態培養基中，對於樟芝菌絲體（mycelia）、葡萄糖胺（glucosamine）與多醣體（polysaccharides）的生產及品質之影響。
試驗結果顯示，以不同濃度的幾丁聚醣為碳源，添加於幾丁聚醣培養基中，與MEA培養基比較。幾丁聚醣培養基之生長速度較緩慢，但菌絲出現明顯的鮮紅色，整體菌絲的生長型態以內部緊實及外緣鬆散方式呈現。於搖瓶培養方面，添加不同去乙醯度的幾丁聚醣（40％、50％、60％）與樟屬抽出物可得到最多葡萄糖胺，分別為25 mg/ml之40％幾丁聚醣（2.25 mM）、15 mg/ml之60％幾丁聚醣（2.24 mM）與牛樟之甲醇抽出物（2.26 mM）；胞外多醣（EPS）分別為50 mg/ml之40％幾丁聚醣（0.212 g）、25 mg/ml之60％幾丁聚醣（0.218 g）與土肉桂之乙酸乙酯抽出物（0.154 g）。隨著添加物濃度增加，胞外多醣的含量也隨之增加。
在抗氧化試驗中，以添加15 mg/ml之40％幾丁聚醣（88.37％）、牛樟之乙酸乙酯抽出物（84.66％）與未添加的培養基皆有較佳的抗氧化力。於捕捉DPPH自由基能力以15 mg/ml之60％幾丁聚醣（78.95％）、牛樟之甲醇抽出物（86.54％）與樟樹之甲醇抽出物（85.97％），其次是未添加幾丁聚醣的菌絲。甲醇萃取濃度在2.5 mg/ml～10 mg/ml時，添加30 mg/ml之40％幾丁聚醣（86.31％）、牛樟之正己烷抽出物（80.19％）與土肉桂之甲醇抽出物（78.19％）表現出較高的螯合亞鐵離子能力。
研究結果顯示，高濃度的幾丁聚醣與樟屬抽出物不僅保有菌絲原本之抗氧化性質，而且能提升其部份抗氧化能力、葡萄糖胺與胞外多醣達到較高的程度。

Taiwanofungus camphorates is an important Chinese medicinal mushroom, and is an unique species only grown in Taiwan. It has many functional properties in human body, especially for promoting immune system and protecting liver. As growing slowly in the wild, supply of T. camphorates with stable product quality at reasonable price is in high demand. Research is interest in attempting to utilize chitosan and Cinnamomum plants extract as substrate for production of this organism, on the productivity and quality of the microbial mycelia, glucosamine and polysaccharides.
Result of the study, as the carbon source with chitosans of different deacetylation degrees, add it in chitosans medium, compare with MEA medium. The mycelium growth rate of chitosans medium is relatively slow in speed, but the brilliant red that the mycelium is obvious, the growth type of the whole mycelium is closely knit with the internal and the loose of rim ppear. Addition of chitosans of different deacetylation degrees (40%, 50%, 60%) and Cinnamomum plants extract can get high content glucosamine in flask cultured, it is 25 mg/ml of 40% chitosans (2.25 mM), 15 mg/ml of 60% chitosans (2.24 mM) and methanol extract of C. kanehirae (2.26 mM), respectively. Part of exopolysaccharides, it is 50 mg/ml of 40% chitosans (0.212 g), 25 mg/ml of 60% chitosans (0.218 g) and ethyl acetate extracts of C. osmophloeum (0.154 g), respectively. As the concentration of additive increases, the content of exopolysaccharides with increase in flask cultured.
In the study of antioxidant activity, in order to addition 15 mg/ml of 40% chitosan (88.37%), ethyl acetate extracts of C. kanehirae (84.66%) and without additive, all has good antioxidant activity in flask cultured. In the study of 1,1-diphenyl-2-picrylhydrazyl radicals. With 15 mg/ml of 60% chitosan (78.95%), methanol extract of C. kanehirae (86.54%) and methanol extract of C. camphora (85.97%), the secondary were mycelium of without chitosan. The mycelium cultured with 40% chitosan of 30 mg/ml (86.31%), n-Hexane extract of C. kanehirae (80.19%) and methanol extract of C. osmophloeum (78.19%) at methanol extract (2.5 mg/ml–10 mg/ml) showed a high chelating effects on ferrous ions.
Summarily, high concentration of chitosan and Cinnamomum plants extract could not only maintain the antioxidant properties of mycelia, but also enhanced some of it is antioxidant properties and higher production of glucosamine and exopolysaccharides.

摘 要 I
ABSTRACT III
誌 謝 V
目　錄 VII
表目錄 X
圖目錄 XI
第1章 緒論 1
1.1 前言 1
1.2 研究背景與動機 1
1.3 研究目的 3
第2章 文獻探討 4
2.1 一般食用藥菇之介紹與生理活性 4
2.2 樟芝之簡介 7
2.2.1 樟芝菌種發展史及型態特徵 7
2.2.2 樟芝成分分析 9
2.3 樟芝的藥理特性與功用 10
2.3.1 葡萄糖胺（Glucosamine）之介紹 10
2.3.2 樟芝多醣（Polysaccharides）之介紹 11
2.3.3 樟芝抗氧化（Antioxidants）性質分析 13
2.4 食用藥菇液態培養之特性 14
2.4.1 培養基組成 15
2.4.2 攪拌速率與剪切力 17
2.4.3 溫度 18
2.4.4 pH值 18
2.4.5 黏度 19
2.4.6 溶氧值（DO值） 19
2.5 幾丁質與幾丁聚醣之介紹 19
2.5.1幾丁質 19
2.5.2幾丁聚醣 22
2.6 添加抽出物對真菌生長型態之作用 24
第3章 材料與方法 25
3.1 試驗流程圖 25
3.1.1 菌絲體培養流程 25
3.1.2 幾丁聚醣備製之流程 26
3.1.3 抽出物萃取之流程 27
3.2 試驗材料 28
3.2.1 樟芝菌株 28
3.2.2 幾丁聚醣 28
3.2.3 樟屬植物 28
3.2.4 培養基 29
3.2.5 試驗藥品與儀器 30
3.3 試驗方法 32
3.3.1 樟芝菌絲體之培育 32
3.3.2 幾丁質與幾丁聚醣之製備 33
3.3.3 樟屬抽出物之萃取與添加 33
3.3.4 樟芝菌絲體固態培養試驗 34
3.3.5 樟芝菌絲體液體震盪培養試驗 35
3.3.6 樟芝菌絲體製備及培養液之分離與純化 36
3.3.7 樟芝培養液之葡萄糖胺測定 36
3.3.8 樟芝培養液之多醣萃取與分析 36
3.3.9 樟芝菌絲體分析抗氧化能力測定 37
3.4 統計分析 38
第4章 結果與討論 39
4.1 固態培養樟芝菌絲體 39
4.1.1 菌絲體於固態培養基之生長速率 39
4.1.2 菌絲體於不同去乙醯度幾丁聚醣下之生長情形 41
4.2 樟芝菌絲體震盪培養之活性生成物 56
4.2.1 不同去乙醯度幾丁聚醣對葡萄糖胺與胞外多醣生成之
影響 56
4.2.2 不同樟屬抽出物對葡萄糖胺與胞外多醣生成之影響 59
4.4 樟芝菌絲體抗氧化能力測定 62
4.4.1 共軛雙烯（Conjugated diene）法測定 62
4.4.2 捕捉1,1-二苯基-2-苦味 基團（DPPH‧）能力測定 68
4.4.3 螯合亞鐵離子能力測定 74
第5章 結論與未來展望 80
5.1 結論 80
5.2 建議 81
參考文獻 82
附錄 90
作者簡介 91

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