臺灣博碩士論文加值系統

人類的平均壽命越來越高，也越來越多人開始注重身體的保養，
目前已有證據顯示自由基與人體疾病的產生有相當程度的關連。有研
究指出動脈粥狀硬化（Witzum,1994），呼吸道疾病（Cross et
al.,1994），神經性退化疾病（Jenner,1994），發炎（Grisham,1994），
癌症（Beckman and Ames,1997），老化（Beckman and Ames,1997）等
疾病的發生都與自由基有關。
樟芝又稱牛樟芝或牛樟菇，只生長在台灣特有的牛樟樹內，屬於
無褶菌目、多孔菌科。在民間盛傳樟芝的功效頗多，而在有關抗氧化
的研究中，Song and Yen（2002）之研究指出樟芝發酵液與發酵菌絲
之萃取物在實驗中具有清除超氧陰離子及抑制脂質過氧化之能力；而
在蔡雁暉（2002）之研究亦明確指出樟芝多醣體具有良好抗氧化能力。
本研究利用氧自由基吸收能力測試（Oxygen-Radical Absorbance
Capacity assay;ORAC assay）､抗脂質過氧化測試（Anti-Lipid
Peroxidiant Capacity assay;ALPC assay）以及總抗氧化（Trolox
Equivalent Antioxidant Capacity;TEAC）測定來探討通氣量對於樟芝醱
酵生產抗氧化物質之影響，並且利用工程的技術使抗氧化物質提升。
由實驗中可以得知以下幾點結果：（1）抗氧化物質與多醣體之生
產是以一次代謝為主。（2）抗氧化能力及多醣體與通氣量
(0.3vvm~1.0vvm)成正比關係。（3）粗多醣在抗脂質過氧化能力與氧
自由基吸收能力中扮演著重要之角色。（4）在各個通氣量
(0.3vvm~1.0vvm) 中，1.0vvm 是抗氧化物質生產之最佳條件。

The human average life span is higher and higher, more and more people too begin maintenance to pay attention to body , have evidence show free radical and human production is it connect with to imprisonning to no small extent of disease already at present. Study and point out scleratheroma (Witzum ,1994),Breathing problem (Cross et al., 1994),(Jenner of disease that the nerve degradates ,1994),Inflammation (Grisham ,1994),Cancer (Beckman and Ames ,1997),Wear out (Beckman and Ames ,1997)Wait for the emergence of the disease to all relate to free radical.
The camphor tree sesame has another name called ox's camphor tree sesame or ox camphor tree mushroom, only grow in peculiar ox camphor tree of Taiwan, have no fungus item of pleat , porous fungus department. A lot of in folk efficiency of widelying spread the camphor tree sesame, in relevant anti-oxidant research, Song and Yen ( 2002) Research point out not having the fermented the sesames liquid of camphor tree it have not lasting oxygen anion not ultra and not suppressing fat the qualities not to pass ability that oxidize of with extraction thing of hypha not fermented on experiment ; Also point out clearly in the research of Cai YanHui (2002 ) that many candy body of camphor tree sesame has good anti-oxidant ability .
This research utilizes oxygen free radical to absorb aptitude tests (Oxygen-Radical Absorbance Capacity assay; ORAC assay), resist fat quality is it oxidize testing to pass (Anti-Lipid Peroxidiant Capacity assay; ALPC assay), and always anti-oxidant (Trolox Equivalent Antioxidant Capacity; TEAC) determine and probe into the keeping in touch with amount to set aside the influence that the ferment produces the anti-oxidant material on the camphor tree sesame, and utilize technology of the project to make the anti-oxidant material improve.
Can learn some following results from the experiment : ( 1) The production of anti-oxidant material and many candy body is to rely mainly on supersession. ( 2) Anti-oxidant ability and much candy body and relation in direct radio of the keeping in touch with amount (0.3vvm resists 1.0vvm ). ( 3) In resist thick many candy fat quality is it oxidize. ( 4) In each keeping in touch with amount (0.3vvm is in 1.0vvm ), 1.0vvm is a optimum condition of the anti-oxidant material production.

摘要…………………………………………………………………… I
目錄……………………………………………………………………Ⅱ
表索引……………………………………………………………..…. ..Ⅶ
圖索引…………..…………………………………………………...…Ⅷ
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…………………………………………………….. 3
2.1 菇類的介紹…………………………………….…………….. 3
2.2 樟芝的介紹…………………………….…………………….. 3
2.3 牛樟芝效用……………………………..……………………. 5
2.4 牛樟芝成分分析……………………….…………………….. 5
2.5 樟芝之藥理作用文獻考… ………………………………….. 6
2.6 抗氧化性質簡介……………………………….…………….. 9
2.6.1 自由基與活性氧之種類…………….…………………….. 9
2.6.2 自由基與活性氧的主要來源……………….…………….. 12
2.6.3 自由基與活性氧的特性……………………..…………….. 14
2.6.4自由基與活性氧分子所引起的氧化傷害………..……….. 18
II
2.6.5自由基與疾病………………………………………..…….. 24
2.6.6 抗氧化劑………………………………………….……….. 24
2.6.6.1 抗氧化劑作用原理及機制…………..………………….. 25
2.6.6.2 天然抗氧化物質…………………………..…………….. 26
2.6.7 抗氧化測試方法…………………………….…………….. 31
2.6.7.1 DPPH(2,2-diphenyl-1-picrylhydrazyl)自由基清除能力測
試……………………………………………...………….. 31
2.6.7.2 還原能力（Reducing power）測試…………………….. 32
2.6.7.3 螯合亞鐵離子（Fe2+）能力測試…………...………….. 32
2.6.7.4 總抗氧化能力（Trolox Equivalent Antioxidant Capacity；
TEAC）測試………………………..………………….. 33
2.6.7.5 氧自由基吸收能力測試（Oxygen-Radical Absorbance
Capacity assay;ORAC assay）…………………...…….. 33
2.6.7.6抗脂質過氧化測試（Anti-Lipid Peroxidiant Capacity
assay;ALPC assay）………………………..………….. 36
2.8 深層液態發酵培養………………………………………….. 35
2.9 影響發酵的物理化學因子………………………………….. 37
2.9.1 培養基組成……………………………….……………….. 37
2.9.2 攪拌速率……………………………….………………….. 40
III
2.9.3 溫度………………………………………….…………….. 41
2.9.4 pH 值……………………………………………………….. 41
2.9.5 黏度(viscosity)…………………………………………….. 41
2.9.6 溶氧值(DO)……………………………………………….. 42
2.9.7 接種量…………………………………………….……….. 42
2.9.8 發酵槽的形式…………………………..………………….. 42
第三章 實驗材料與方法…………………………………………….. 44
3.1 實驗材料…………………..……………………………….. 44
3.1.1 實驗菌株………………………………………………….. 44
3.1.2 實驗藥品………………………………………………….. 44
3.1.3 實驗儀器及其他設備…………………………………….. 45
3.1.4 實驗裝置………………………………………………….. 46
3.2 實驗方法…………………………………………………….. 47
3.2.1 菌株保存………………….……………………………….. 47
3.2.2 培養基組成……………………………………………….. 47
3.2.3 操作條件………………………………………………….. 48
3.2.4 分析方法……………………….………………………….. 49
3.2.4.1 發酵液處理流程………….…………………………….. 49
3.2.4.2 菌重分析…………………...…………………………….. 49
IV
3.2.4.3 葡萄糖殘量分析(還原醣測定)………………………….. 50
3.2.4.4 多醣體分析…………………………………………….... 50
3.2.4.4.1 粗多醣之分離………………………………………….. 50
3.2.4.4.2 酚-硫酸呈色法分析(總多醣)………………………….. 51
3.2.4.4.3 β-(1-3) glucan 含量測定-螢光結構活性測定法…..….. 52
3.2.4.5 抗氧化能力分析…………….………………………….. 53
3.2.4.5.1 氧自由基吸收能力分析（Oxygen-radical absorbance
capacity assay;ORAC assay）……………………….. 53
3.2.4.5.2 總抗氧化能力（Trolox equivalent antioxidant capacity
assay;TEAC assay）………………………………….. 54
3.2.4.5.3 抗脂質過氧化測試…………………..……………….. 55
第四章 結果與討論………………………………………………….. 56
4.1 各個測試方法之標準曲線………………………………….. 56
4.1.1 葡萄糖殘量分析之標準曲線……………………….…….. 56
4.1.2 酚-硫酸呈色法分析之標準曲線………..…………….….. 56
4.1.3 氧自由基吸收能力分析之標準曲線…………….……….. 57
4.1.4總抗氧化能力能力分析之標準曲線……………..………. 57
4.2 樟芝發酵液中抗氧化物質之代謝趨勢…………………….. 58
4.3 在不同通氣量下各種抗氧化能力之變化………………….. 63
V
4.3.1 在不同通氣量下氧自由基吸收能力之變化………..…….. 63
4.3.2 在不同通氣量下抗脂質過氧化之變化………………….. 64
4.3.3 在不同通氣量下總抗氧化能力之變化………………….. 65
4.3.4 在不同通氣量下β-D-glucan 含量之變化……………….. 66
4.4 在不同通氣量下β-D-glucan 對於各種抗氧化能力之影響.. 67
4.4.1 在各通氣量下β-D-glucan 對於氧自由基吸收能力分析之影
響……………………………………………………….…..67
4.4.2 在各通氣量下β-D-glucan對於抗脂質過氧化能力測試之影
響……………………………………………………….….. 68
4.4.3 在各通氣量下β-D-glucan對於總抗氧化能力之影響….. 69
4.5 選擇最佳生產條件………………………………………….. 70
4.5.1 產生菌體之最佳條件…………………………….……….. 70
4.4.2 產生產物之最佳條件…………………………….………..71
第五章 結論與建議………………………………………………….. 77
5-1 結論………………………………………………………..... 77
5-2 建議…………………………….……………………..…….. 77
參考文獻………………………………………………………………. 79

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