臺灣博碩士論文加值系統

酒粕是酒醪經壓榨後所形成的粕，也是釀酒中所形成之廢棄物，科學家在釀酒廠中偶然發現製酒工人的手總是白皙光滑，使得人們開始著手進行研究。而目前研究大多數皆為清酒酒粕，對於水果酒粕的研究卻是少之又少。本篇論文主要利用台灣特有種真菌牛樟芝與桑椹、山藥及葡萄酒粕進行發酵，測定其吸收紫外線、抑制黑色素及Tyrosinase活性，期望藉由發酵來提高酒粕的功能性與應用價值，使酒粕從廢棄物轉變為市場上的商機。

實驗結果顯示，在吸收紫外線實驗中，只有山藥酒粕乙醇萃取液在280nm下具有吸收紫外線效果。抑制Tyrosinase體外實驗顯示，三種未經過發酵酒粕皆無抑制Tyrosinase功效，經過牛樟芝與酒粕合併發酵後，其酒粕菌絲體水萃取液具有抑制Tyrosinase功效，抑制效果為山藥酒粕>桑椹酒粕>葡萄酒粕，酒粕菌絲體乙醇萃取液皆無抑制Tyrosinase，而發酵液結果顯示只有山藥酒粕具有抑制Tyrosinase效果。以細胞實驗顯示，酒粕能抑制細胞中的Tyrosinase，且亦能透過牛樟芝發酵來提高抑制Tyrosinase，酒粕菌絲體水萃取液抑制Tyrosinase效果為桑椹酒粕>山藥酒粕>葡萄酒粕，山藥酒粕發酵液對Tyrosinase抑制能力在50ppm濃度下為63.7±0.6%。最後再經由SDS-PAGE實驗發現，酒粕萃取液及發酵液確實可以抑制Tyrosinase活性，但無法抑制Tyrosinase表現量。

Wine lees is unfiltered by the press after the formation of the lees, wine is also formed in the waste, 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. In This study use yam, grapes, and mulberry wine lees were used as substrates, and fermented by Antrodia cinnamomea., measured by UV absorption, inhibition of melanin and Tyrosinase activity, expected to improve the function and value of wine lees by fermented, the wine lees from the waste to market opportunities.

The experimental results show that in the absorption of ultraviolet radiation experiments, only the ethanol extract of yam wine lees has the absorption of ultraviolet light under the effect of 280nm. Inhibition of Tyrosinase in vitro experiments showed that the three wine lees has not been fermented showed no inhibition of Tyrosinase, but wine lees by Antrodia cinnamomea co-fermented , the wine lees mycelium water extract can inhibit Tyrosinase, inhibitory effect of the yam wine lees> mulberry wine lees> grapes wine, ethanol extract of mycelium wine lees showed no inhibition of Tyrosinase, the fermentation broth revealed that only yam wine lees can inhibition of Tyrosinase. To cell experiments show that the wine lees can inhibit the Tyrosinase, and also by Antrodia cinnamomea co-fermented to increase the inhibition of Tyrosinase, dregs mycelium water extract inhibited Tyrosinase of mulberry wine lees> yam wine lees> grapes wine lees, yam wine lees fermentation broth Tyrosinase inhibition of fermentation liquid body the ability to 50ppm concentration was 63.7 ± 0.6%. Finally, was found by SDS-PAGE, ferment broth and mycelia extract of wine lees can actually inhibit the Tyrosinase activity, but did not inhibit Tyrosinase expression.

摘要....................................................................................................................................I
Abstract.............................................................................................................................II
目次.................................................................................................................................IV
圖目錄............................................................................................................................VII
表目錄.............................................................................................................................IX
第一章 緒論.....................................................................................................................1
1.1 前言....................................................................................................................1
1.2 研究動機與目的................................................................................................1
第二章 文獻回顧.............................................................................................................3
1. 酒粕......................................................................................................................3
1.1 酒粕簡介......................................................................................................3
1.2 酒粕成分......................................................................................................3
1.3 酒粕功效......................................................................................................3
2. 牛樟芝..................................................................................................................4
2.1 牛樟芝簡介..................................................................................................4
2.2 牛樟芝的成分..............................................................................................5
2.3 牛樟芝的功效..............................................................................................6
3. 桑椹......................................................................................................................7
3.1 桑椹簡介......................................................................................................7
3.2 桑椹的成分..................................................................................................8
3.3 桑椹的功效..................................................................................................8
4. 山藥....................................................................................................................8
4.1山藥簡介.......................................................................................................8
4.2 山藥的成分..................................................................................................8
4.3 山藥的功效................................................................................................9
5. 葡萄................................................................................................................9
5.1葡萄簡介.....................................................................................................9
5.2 葡萄成分..................................................................................................10
5.3 葡萄酒成分與功效..................................................................................10
6. 皮膚結構......................................................................................................11
6.1 表皮..........................................................................................................11
6.2 真皮層......................................................................................................12
6.3 皮下組織..................................................................................................13
7. 紫外線..........................................................................................................13
7.1 紫外線的分類..........................................................................................13
7.2 紫外線對人體的傷害..............................................................................15
8. 防曬原理......................................................................................................15
8.1 化學性防曬劑..........................................................................................16
8.2 物理性防曬劑..........................................................................................16
9. 黑色素..........................................................................................................17
9.1 黑色素體結構與功能..............................................................................17
9.2 黑色素形成..............................................................................................18
9.3 酪胺酸酶活性的抑制..............................................................................19
9.4 防止或抑制黑色素形成之機制..............................................................20
9.5 市售美白成分抑制黑色素機制..............................................................20
第三章 材料與方法.......................................................................................................23
3.1 實驗架構....................................................................................................23
3.2 實驗材料....................................................................................................24
3.3 實驗藥品....................................................................................................24
3.4 儀器設備....................................................................................................25
3.5 實驗方法....................................................................................................25
3.5.1 酒粕萃取液製備...................................................................................25
3.5.2 樟芝菌絲體液態發酵...........................................................................25
3.5.3 萃取液及發酵液之吸光值測定...........................................................26
3.5.4 Tyrosinase抑制能力測試......................................................................26
3.5.5 細胞培養...............................................................................................27
3.5.6 萃取液及發酵液對細胞存活率之影響...............................................28
3.5.7 Tyrosine hydroxylase活性試驗.............................................................29
3.5.8蛋白質定量試驗....................................................................................29
3.5.9 SDS-PAGE 蛋白質電泳試驗...............................................................30
第四章 結果與討論.......................................................................................................32
4.1牛樟芝生長曲線.........................................................................................32
4.2各種酒粕發酵前後之UV-A、UV-B、UV-C吸收值變化......................33
4.3 抑制Tyrosinase活性測定..........................................................................38
4.4 酒粕與牛樟芝發酵產物對老鼠黑色素瘤細胞(B16-F1)之影響.............44
4.5 Tyrosinase hydroxylase活性分析..............................................................49
4.6 以SDS-PAGE探討各種酒粕萃取液及發酵液對老鼠黑色素瘤細胞
Tyrosinase表現量之影響..........................................................................55
第五章 結論...................................................................................................................62
第六章 參考文獻...........................................................................................................64

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