臺灣博碩士論文加值系統

樟芝(Antrodia camphorata)只寄生於台灣原生種之牛樟樹的中空腐朽樹幹內。本研究先透過掃描式電子顯微鏡確認樟芝菌種，並作為研究材料，將樟芝菌絲體培養於不同固液態培養基中並以4,7-dimethoxy-5- methyl-1,3-benzodioxole(DMB)及4-acetylantroquinonol B(4B)兩種成分為指標，進行不同菌種、不同培養基及不同採收時期發酵培養產物之HPLC成分分析，同時針對樟芝子實體及不同培養基與不同採收時期之菌絲體，測試抑制酪胺酸酶、DPPH自由基清除能力、還原能力等體外抗氧化活性試驗。在成分定量分析獲得DMB於固態培養菌絲體比液態培養菌絲體之含量較高，以紅薏仁為培養基其含量最高為7579.4± 95.6 μg/g D.W.；而4 B則以台梗九號培養樟芝菌絲體為最多含量為3182.6 ±159.4 μg/g D.W.，比其他固態培養基高出5倍多，這兩種化合物於樟芝子實體中含量皆偏低。另外，抑制酪胺酸酶之活性，以75% ethanol萃取之萃取物於1000 μg/ mL時，以樟芝子實體抑制酪胺酸酶為最高，其抑制率為50.4± 2.3 %；體外抗氧化試驗中，以100% methanol萃取之紅薏仁培養菌絲體，擁有最佳之清除DPPH自由基之能力，其SC50為148.7 μg/mL，另外還原力試驗方面也顯示以紅薏仁培養菌絲體有較高之還原能力。
關鍵字：樟芝(Antrodia camphorata)、DPPH清除能力、DMB、4B、高效液相層析儀

Antrodia camphorata parasitized only in the inner cavity of the rotten trunk of Cinnamonum kanehirae Hay., an important endemic tree species in Taiwan. In this study, A. camphorata mycelial strains were identified using scanning electronic microscope (SEM), and, then, 4,7-dimethoxy-5-methyl- 1,3-benzodioxol and 4-acetylantroquinonol B were used as two marker components to investigate the components in the fermented products of different A. camphorata mycelia strains grown on different culturing media in different harvest time using high performance liquid chromatography (HPLC). Meanwhile, the antioxidant bioactivities including inhibition of tyrosinase, DPPH free radical scavenging ability, and reducing ability were tested in vitro using A. camphorata fruit bodies which were grown on different culturing media and in different harvested time. Results showed that A. camphorata mycelia which were grown on solid substrates, especially those grown on red coix had the highest content of 4,7-dimethoxy-5-methyl-1,3-benzodioxol (7579.4± 95.6 μg/g dry weight (D.W.)). Meanwhile, A. camphorata mycelia which were grown on Taiken 9 brown rice had highest 4-acetylantroquinonol B content (3182.6± 159.4 μg/g D.W.) and the content was of more than 5 times than those harvested from other culturing substrates. However, both of the two components were low in fruit bodies of A. camphorata. The 75% ethanol extract of fruit bodies of A. camphorata had the highest inhibition ability of tyrosinase (50.4± 2.3%) when the dose was at 1000 μg/mL. The 100% methanol extract of the mycelia harvested from red coix had highest DPPH free radical scavenging ability and SC50 was at 148.7 μg/mL. Moreover, the mycelia harvested from red coix had the highest reducing ability.

中文摘要.................................................... Ⅰ
Abstract.................................................. Ⅱ
致謝...................................................... Ⅳ
目錄...................................................... Ⅴ
圖表目錄.................................................. Ⅷ
壹、前言................................................... 1
貳、文獻回顧................................................ 3
一、牛樟樹的簡介............................................. 3
二、樟芝的介紹.............................................. 4
三、樟芝培養之介紹........................................... 7
四、樟芝之組成成分.......................................... 10
五、酪胺酸酶(Tyrosinase)與酪胺酸酶抑制物...................... 14
六、樟芝之抗氧化............................................ 16
參、材料與方法............................................. 18
一、實驗架構............................................... 18
二、試驗材料............................................... 20
三、試驗方法............................................... 23
(一)培養樟芝菌絲........................................... 23
(二)菌種鑑定.............................................. 24
(三)樟芝固態培養條件之試驗................................... 26
(四)樟芝液態菌絲體之試驗..................................... 27
(五) 樟芝發酵產物萃出液生物活性物質之定量分析試驗 ............... 28
(六)樟芝菌絲體之酪胺酸酶(Tyrosinase)之活性分析................. 32
(七)樟芝體外抗氧化活性試驗................................... 33
(八)統計分析.............................................. 34
肆、結果.................................................. 35
一、樟芝菌種鑑定............................................ 35
二、樟芝(A. camphorata)菌絲體固液態培養...................... 37
三、樟芝成分之定量分析....................................... 43
四、樟芝(A. camphorata)發酵產物定量分析試驗................... 49
五、樟芝菌絲體之酪胺酸酶(Tyrosinase)活性分析與體外抗氧化之評估.... 59
伍、討論.................................................. 68
一、樟芝菌種鑑定............................................ 68
二、樟芝固液態培養產物生長條件之探討........................... 68
三、成分定量分析............................................ 70
四、樟芝菌絲體之酪胺酸酶(Tyrosinase)活性分析與體外抗氧化之活性....................................................... 72
陸、結論.................................................. 74
參考文獻.................................................. 75
附錄..................................................... 86
作者簡介.................................................. 89

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