探討添加界面活性劑Tween 80 對Penicillium brevicompactum 在液態發酵中生產Mycophenolic acid 之影響

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羅心怡(Hsin-Yi Lo) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

探討添加界面活性劑Tween 80 對Penicillium brevicompactum 在液態發酵中生產Mycophenolic acid 之影響
(Effects of surfactant Tween 80 on the production of Mycophenolic acid by Penicillium brevicompactum in batch culture)

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摘要(中)

自器官移植的歷史揭開序幕之後，如何降低排斥的現象，一直是醫界致力解決的問題，因應而生的即是抗排斥的藥物。本研究中的Mycophenolic acid (MPA)是用來作為器官移植時抑制免疫反應的藥物，作為Inositol MonoPhosphate DeHydrogenase (INPDH)之非競爭型抑制劑，針對淋巴細胞之合成路徑進行抑制而達到降低免疫反應的發生，而實驗所使用微生物是Penicillium brevicompactum進行深層液態發酵(submerged fermentation)生產MPA。
　　為了提高P. brevicompactum生產Mycophenolic acid的產量，本研究採用深層液態發酵的方式，以添加界面活性劑Tween 80作為變因探討Mycophenolic acid的產量，並進一步了解發酵過程中界面活性劑Tween 80扮演的角色及其對生產Mycophenolic acid的影響。
　　整體而言添加界面活性劑Tween 80對於P. brevicompactum生產Mycophenolic acid是有正面幫助。本研究得到的結論為當添加界面活性劑Tween 80為0.2%時得到最佳Mycophenolic acid產量為0.512 g/l，提升1.6倍之多；Productivity提升2.1倍為4.196 mg/hr。藉由KLa及SOUR 推測Tween 80的添加扮演促進氧氣傳送的角色，且添加Tween 80也會改變其P. brevicompactum的菌體的型態，又以有緻密核心且外圍毛茸鬆散狀的菌球有最佳的MPA產量。

摘要(英)

Mycophenolic acid (MPA) is a organic acid and had been discovered as early as 1983 by an Italian physician. MPA against to bacteria, fungi, viruses, toumours, psoriasis, and inhibit the grorth of the anthrax bacillus. Mycophenolate mofetil (MMF) is a prodrug of mycophenolic acid, an inhibitor of inosin monophosphate dehydrogenase (IMPDH).Mycophenolate mofetil (MMF), an ester of mycophenolic acid, has been widely used as an immunosuppressant in kidney, heart, and liver transplantationand as a medication to treat other ailments.
Oxygen transport rates limit the productivites of aerobic fermentations. Penicillium brevicompactum is an aerobic microorganism and improve cell growth by supplying sufficient oxygen. In this study, dissolved oxygen in the fermentation medium plays an important environment factor to affect not only cell growth but also MPA concentration. As the air flow rate was controlled as 0.2vvm、0.5vvm of each batch ,the maximum MPA concentration increased from 0.324 g/l to 0.462 g/l ; Product yield rate increased from25.9 mg/g to 36.9 mg/g. Tween80 addition had shown to increase cell growth and MPA production. The highest MPA production, 0.512 g/l, were obtained when 0.2 % Tween80 were added, which was 1.6-fold of the control. The productivity increased 2.1-fold of the control. Surfact-active substances affect pellet growth. With Penicillium brevicompactum, the large pellets develop with the outer hairy region of pellets when Tween80 was added.

關鍵字(中)

★ 青黴菌
★ 麥考酚酸
★ 界面活性劑

關鍵字(英)

★ Mycophenolic acid
★ Tween 80
★ Penicillium brevicompactum

論文目次

摘要 I
目錄 II
圖索引 V
表索引 IX
第一章緒論 1
1.1研究動機 1
1.2研究目的 2
第二章文獻回顧 4
2.1應用微生物之發展 4
2.1.1真菌介紹 4
2.1.2真菌的分類 5
2.1.3青黴菌之介紹 6
2.1.4藥用抗生素的時代 7
2.2 Mycophenolic acid之介紹 10
2.2.1Mycophenolic acid 之發現 10
2.2.2 Mycophenolic acid之生化合成 10
2.2.3 Mycophenolic acid在醫療上之應用 11
2.3人體免疫系統和器官移植之簡介 13
2.3.1人體免疫系統簡介 13
2.3.2器官移植簡介 14
2.3.3器官移植免疫學 15
2.3.4器官排斥 17
2.3.5人體器官移植時所使用的藥物 19
2.3.6 Mycophenolic acid應用在器官移值上 24
2.4發酵動態行為的數學模式 27
2.4.1與生長有關的產物(growth-associated product) 27
2.4.2與生長無關的產物(nongrowth-associated product) 27
2.4.3混合生長相關的產物(mixed-growth-associated product) 28
2.5氧氣對微生物的影響 29
2.5.1溶氧值(DO值) 29
2.5.2溶氧值對於青黴菌之影響 30
2.5.3氧氣質傳對於菌體生長與產物生成之影響 31
2.5.4氧氣質傳係數之影響因子 34
2.5.4.1攪拌速率 35
2.5.4.2曝氣速率 36
2.5.4.3額外添加物 37
2.6界面活性劑 39
2.6.1界面活性劑的分類 39
2.6.2界面活性劑在生物程序之應用 40
第三章實驗規劃、材料與方法 45
3.1實驗規劃 45
3.2 實驗材料 46
3.2.1 實驗藥品 46
3.2.2實驗儀器及其他設備 48
3.2.3 實驗裝置 49
3.3 實驗菌株與方法 50
3.3.1實驗菌株 50
3.3.2 培養基組成 50
3.4實驗方法 52
3.4.1菌種接種及保存方法 52
3.4.2發酵培養 53
3.4.2.1不同供氧量之震盪培養實驗 53
3.4.2.2添加不同種氧氣因子之震盪培養實驗 53
3.4.2.3在不同供氧量下添加不同濃度Tween 80之震盪培養實驗 54
2.4.2.4液態發酵改變不同的通氣量之發酵槽實驗 54
2.4.2.5液態發酵添加不同濃度Tween 80之發酵實驗 55
3.4.3實驗分析方法： 56
3.4.3.1發酵液處理流程 56
3.4.3.2菌重分析 57
3.4.3.3殘糖分析(Sucrose)-酚-硫酸呈色法分析 58
3.4.3.4產物濃度(MPA)之分析 59
第四章實驗結果與討論 61
4.1 震盪培養實驗 61
4.1.1不同通供氧量對Penicillium brevicompactum生產Mycophenolic acid影響 61
4.1.2不同種氧氣因子對Penicillium brevicompactum生產Mycophenolic acid的影響 62
4.1.2.1不同界面活性劑對對Penicillium brevicompactum生產MPA之影響 62
4.1.2.2添加n-hexadecane對Penicillium brevicompactum生產MPA之影響 64
4.1.3在不同工作體積下添加不同濃度界面活性劑Tween80之影響 67
4.2 發酵槽培養實驗 70
4.2.1發酵槽實驗之通氣量(aeration rate)對Penicillium brevicompactum 生產MPA之影響 71
4.2.1.1發酵動力曲線的比較 71
4.2.1.2 SOUR值對於菌體生長的影響 74
4.2.1.3 SOUR值對於產物MPA的影響 75
4.2.2 發酵槽實驗之通氣量0.2 vvm下添加Tween80對Penicillium brevicompactum 生產MPA之影響 79
4.2.2.1發酵動力曲線的比較 79
4.2.2.2添加Tween 80對於菌體生長的影響 82
4.2.2.3 添加Tween 80對於產物MPA的影響 83
4.3菌體型態(morphology)對Penicillium brevicompactum生產
Mycophenolic acid 的影響 88
4.3.1不同通氣量對Penicillium brevicompactum生產 Mycophenolic acid之morphology的影響 88
4.3.2添加Tween 80對Penicillium brevicompactum生產Mycophenolic acid之morphology的影響 91
第五章結論與建議 94
5.1結論 94
5.2建議 95
第六章參考文獻 96

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指導教授

徐敬衡(Chin-hang Shu)

審核日期

2008-7-17

推文