臺灣博碩士論文加值系統

酒粕為釀酒產業中主要的廢棄物，首次被注重是因為科學家偶然發現製酒工人的手總是白皙，才開始著手研究。對於酒粕的研究及應用性皆以清酒粕為主，鮮少以水果酒粕為研究對象。本篇論文利用台灣特有真菌牛樟芝分別與山藥、葡萄及桑椹酒粕萃取液進行液態發酵，測定不同酒粕萃取液與樟芝合併發酵後之發酵產物，其樟芝菌絲體產量、抗氧化功能性評估及細胞氧化傷害保護試驗，期望以此方式能提高水果酒粕的再利用性及樟芝保健抗氧化機能性，並且期待能提供一個酒粕應用上的新方向。
實驗結果顯示，樟芝與2%桑椹酒粕乙醇萃取液合併發酵，可將樟芝菌絲體產量由6.19g/L提至9.02g/L、樟芝菌絲體乙醇萃取液的DPPH清除能力提高17.86%、並且將萃取液總多酚含量由38.48ppm提高至62.16ppm。樟芝與葡萄酒粕水萃取液合併發酵可提高發酵液還原能力，與乙醇萃取液發酵可將菌絲體乙醇萃取液螯合亞鐵離子能力提升27.97％。而樟芝與山藥酒粕水萃取液合併發酵之發酵液螯合亞鐵離子能力提高25.69%，並且提升發酵液清除過氧化氫的能力。最後在細胞氧化傷害保護試驗中，樟芝與桑椹酒粕水萃取液、乙醇萃取液，以及葡萄、山藥酒粕乙醇萃取液合併發酵後，其樟芝菌絲體皆具有提升細胞抗氧化傷害的能力。

Wine lees is the main offal for wine-making industry. Scientist began to investigate the wine lees after they noticed that hands of the wine-making workers are whiter than those of others. The main research and application of wine lees were focused on Sake lees, but rarely on fruit wine lees. This study used yam, grapes, and mulberry wine lees extracts were used as substrates, and fermented by Antrodia camphorata. To investigate the influences on mycelia growth as well as on the antioxidant properties and the H2O2-induced oxidation damage tolerance of the fermentation products. The results obtained in this study is expected to diversify the potential applications of various wine lees fermented by medical fungi.
The experimental results showed that Antrodia camphorata co-fermented with 2% of mulberry wine lees ethanol extracts can improve mycelia production from 6.19g / L to 9.02g / L. The DPPH scavenging ability of mycelia was enhanced by 17.86%, and the contents of total phenols of mycelia was increased from 38.48 ppm to 62.16 ppm. Antrodia camphorata co-fermented with the water extracts of grapes wine lees can enhance the reducing power of fermentation broth. The ethanol extracts of grapes lees can raise the chelating effects of Antrodia camphorata mycelia on ferrous ions by 27.97%. Antrodia camphorata co-fermented with water extract of yam wine lees can improve 25.69% of chelating capability on ferrous ions, and the H2O2 scavenging ability of fermentation broth was also enhanced as compared with control. Finally, in the H2O2-induced oxidation damage test, the anti-oxidative damage ability of the cells can be enhanced by the Antrodia camphorata mycelia co-fermented with water extracts of mulberry wine lees, ethanol extracts of mulberry wine lees, grapes wine lees, and yam wine lees.

目次..............................................................................................................................Ⅰ
圖目錄.........................................................................................................................III
表目錄.........................................................................................................................VI
摘要................................................................................................................................1
Abstract.........................................................................................................................2
第一章 緒論..................................................................................................................3
1.1前言....................................................................................................................3
1.2研究動機及目的................................................................................................4
第二章 文獻回顧..........................................................................................................5
2.1樟芝....................................................................................................................5
2.1.1樟芝簡介...................................................................................................5
2.1.2樟芝成分...................................................................................................6
2.1.2樟芝之藥理作用.......................................................................................6
2.2山藥....................................................................................................................8
2.2.1山藥簡介...................................................................................................8
2.2.2山藥成分...................................................................................................8
2.2.2山藥之藥理作用.......................................................................................9
2.3葡萄..................................................................................................................10
2.3.1葡萄簡介.................................................................................................10
2.3.2葡萄成分.................................................................................................11
2.3.2葡萄之生理活性.....................................................................................11
2.4桑椹簡介..........................................................................................................12
2.5自由基與氧化壓力之介紹..............................................................................12
2.5.1自由基定義.............................................................................................12
2.5.2活性氧定義.............................................................................................13
2.5.3氧化壓力之傷害.....................................................................................16
2.5.4抗氧化防禦系統.....................................................................................20
第三章 實驗架構........................................................................................................23
第四章 研究方法........................................................................................................24
4.1實驗材料..........................................................................................................24
4.1.1酒粕樣品、實驗菌及細胞株.................................................................24
4.1.2實驗藥品.................................................................................................24
4.1.3實驗設備.................................................................................................25
4.2實驗方法..........................................................................................................25
4.2.1酒粕萃取液製備.....................................................................................25
4.2.2液態發酵培養.........................................................................................25
4.2.3抗氧化能力評估.....................................................................................26
4.2.3.1清除α，α-diphenyl-β-picrylhydrazyl(DPPH)...........................26
4.2.3.2還原能力測試................................................................................27
4.2.3.3總多酚含量....................................................................................28
4.2.3.4螯合亞鐵離子................................................................................29
4.2.3.5清除過氧化氫................................................................................30
4.2.4細胞測試.................................................................................................30
4.2.4.1細胞培養........................................................................................30
4.2.4.2 MTT...............................................................................................30
4.2.5統計分析.................................................................................................31
第五章 結果與討論....................................................................................................32
5.1樟芝液態發酵生長曲線測定..........................................................................32
5.2酒粕對樟芝菌絲體生長影響測定..................................................................34
5.3清除α，α-diphenyl-β-picrylhydrazyl(DPPH)................................................36
5.4清除過氧化氫能力測定..................................................................................43
5.5還原能力測試..................................................................................................49
5.6螯合亞鐵離子能力測定..................................................................................55
5.7總多酚含量測定..............................................................................................61
5.8酒粕與牛樟芝發酵產物對老鼠巨噬細胞(RAW 264.7)之影響....................67
5.9細胞氧化傷害保護試驗..................................................................................83
第六章 結論................................................................................................................99
參考文獻....................................................................................................................100

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