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研究生:陳宏文
研究生(外文):Hung-Wen Chen
論文名稱:以回應曲面法探討樟芝高密度發酵並分析相關的成份與機能性
論文名稱(外文):Studies on the High Density Culture of Anthodia cinnamomea with Response Surface Methodology
指導教授:何小台
指導教授(外文):Chester S. Ho
學位類別:碩士
校院名稱:國立交通大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:92
中文關鍵詞:樟芝高密度發酵多醣回應曲面法
外文關鍵詞:Antrodia cinnamomeaHigh cell density fermentationPolysaccharideResponse surface methodologyRSM
相關次數:
  • 被引用被引用:27
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  • 收藏至我的研究室書目清單書目收藏:1
「樟芝」(Antrodia cinnamomea)是極為昂貴的藥用真菌,又稱「牛樟芝」、「牛樟菇」、「樟菇」、「紅樟菰」等。自然界中,發現樟芝僅生長於樟科的牛樟樹(Cinnamomum kanehirai Hay),牛樟樹屬於台灣稀有保育樹種,也因此,天然生長的樟芝亦不多。所以本論文的目的是以高密度發酵的方式,大量生產樟芝菌絲體,並瞭解在培養環境改變的情況下,對相關產物、機能性的影響。
研究的試驗菌株是Antrodia cinnamomea CCRC35396與CCRC35398,以傳統的一次一因子法配合回應曲面法(RSM),設計並分析樟芝高密度發酵的最適培養條件。先運用一階設計中的25-1部份因子設計法,將樟芝以搖瓶震盪實驗,探討培養液中碳源的濃度、氮源的濃度、無機鹽添加與起始pH值等因子,對樟芝菌絲體生長、發酵液終點pH值、多醣生成量、抗氧化力的影響;再利用陡升路徑法、中心混成及二階設計,逐步定出最佳培養條件。
對Antrodia cinnamomea CCRC35398而言,在500 mL有溝槽的搖瓶培養中,裝100 mL培養液,以溫度28 ℃、轉速50 r.p.m.、接種量10 ﹪的固定條件下,先以一次一因子法固定起始pH值為5,然後再由回應曲面法可得最佳培養環境為Glucose 3 ﹪、Sucrose 2 ﹪、Peptone 1.3 ﹪、Yeast extract 1.5 ﹪、Ion 0.03 ﹪。跟原培養條件相比,菌絲體產量已經有顯著的提高,由此可知,在尋找樟芝的最適生長條件上,回應曲面法具有良好的契合度。
Antrodia cinnamomea is precious and rare fungus using for Chinese medicine. In the natural world, they are only found in Cinnamomum kanehirai Hay that protecting in Taiwan. The purpose of study is going to enhance production of Mycelium and analyze the Polysaccharide concentration and antioxidation capacity in different conditions.
The strategy is to find out the optimum cultured condition of Antrodia cinnamomea CCRC35396 and CCRC35398 with one-factor-at-a-time method and experimental design (response surface methodology). Screening for carbon sources and nitrogen sources are tested by one-factor-at-a-time method. Then we can determine the concentration of nutrients by response surface methodology (RSM). RSM is a systemic manner of statistic and including three steps: the 25-1 fractional factorial of first order, the path of steepest ascent and the central composite design of second order.
The optimum cultured conditions of Antrodia cinnamomea CCRC35398 are Glucose 3 %, Sucrose 2 %, Peptone 1.3 %, Yeast extract 1.5 %, Ion 0.03 % when culture in 500 mL Hinton flask containing 100 mL media, 28 ℃, 50 r.p.m., initial pH 5. Results of Mycelium concentrate and Polysaccharide concentrate are increased significantly.
標號標題頁碼
中文摘要i
英文摘要ii
誌謝iii
總目錄iv
附錄實驗目的與策略vii
壹序言簡介1
貳文獻回顧5
參研究方法26
肆實驗結果與討論35
表4-1:點滴培養觀察記錄36
圖4-1:樟芝的固態與液態培養形態37
表4-2:液態培養觀察記錄38
表4-3:菌種35396的碳源篩選40
圖4-2:不同種類碳源對菌種35396菌絲體濃度與終點pH值的影響40
圖4-3:不同種類碳源對菌種35396培養濾液中多醣濃度的影響41
表4-4:菌種35398的碳源篩選42
圖4-4:不同種類碳源對菌種35398菌絲體濃度與終點pH值的影響42
圖4-5:不同種類碳源對菌種35398培養濾液中多醣濃度的影響43
表4-5:菌種35396的氮源篩選44
圖4-6:不同種類氮源對菌種35396菌絲體濃度與終點pH值的影響44
圖4-7:不同種類氮源對菌種35396培養濾液中多醣濃度的影響45
表4-6:菌種35398的氮源篩選46
圖4-8:不同種類氮源對菌種35398菌絲體濃度與終點pH值的影響46
圖4-9:不同種類氮源對菌種35398培養濾液中多醣濃度的影響47
表4-7:一階設計,25-1部份因子設計49
表4-8:一階設計之組別與菌乾重結果50
表4-9:一階設計之碳/氮比與菌乾重結果51
圖4-10:不同C/N比對菌絲體濃度之散佈圖52
表4-10:一階設計之組別與多醣量53
表4-11:一階設計之組別與終點pH值結果54
圖4-11:一階設計中,各組別的菌乾重與終點pH值55
表4-12:一階設計之組別與捕捉DPPH結果56
表4-13:一階設計之組別與以硫氰酸鐵法測得結果57
表4-14:最陡升設計58
表4-15:最陡升設計的菌乾重與終點pH值59
圖4-12:最陡升菌乾重圖59
表4-16:二階設計層階60
表4-17:二階設計61
表4-18:培養基配置62
表4-19:二階組別與菌乾重結果63
表4-20:二階組別與多醣量結果64
表4-21:二階菌乾重統計分析結果65
表4-22:二階多醣量統計分析結果65
圖4-13:Peptone與Yeast extract對菌乾重的曲面圖66
圖4-14:Peptone與Yeast extract對菌乾重的等高線圖66
圖4-15:Glucose與Yeast extract對菌乾重的曲面圖67
圖4-16:Glucose與Yeast extract對菌乾重的等高線圖67
圖4-17:Glucose與Peptone對菌乾重的二階曲面圖68
圖4-18:Glucose與Peptone對菌乾重的等高線圖68
圖4-19:Glucose與Peptone對多醣的二階曲面圖69
圖4-20:Glucose與Peptone對多醣的等高線圖69
圖4-21:Glucose與Yeast extract對多醣的曲面圖70
圖4-22:Glucose與Yeast extract對多醣的等高線圖70
圖4-23:Peptone與Yeast extract對多醣的曲面圖71
圖4-24:Peptone與Yeast extract對多醣的等高線圖71
表4-23:二階培養之碳/氮比對菌乾重的影響72
圖4-25:二階培養之碳/氮比對菌乾重的影響73
表4-24:起始pH值對菌種35396之影響74
圖4-26:起始pH值對菌種35396菌體濃度與終點pH值之影響75
圖4-27:起始pH值對菌種35396培養濾液多醣量之影響75
表4-25:起始pH值對菌種35398之影響76
圖4-28:起始pH值對菌種35398菌體濃度與終點pH值之影響76
圖4-29:起始pH值對菌種35398培養濾液多醣量之影響77
圖4-30:本實驗所使用的500 mL有擋板震盪搖瓶與一般實驗用搖瓶77
伍結論78
陸參考文獻80
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