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研究生:薛煜彬
研究生(外文):Yu-Pin Hsueh
論文名稱:葡萄與葡萄細胞乙醇萃取物對抑制酪胺酸酶活性之效能評估
論文名稱(外文):The inhibitory effects of ethanol extracts from fresh grape and grape cell on mushroom tyrosinase activity
指導教授:許成光
指導教授(外文):Cheng-Kuang Hsu
學位類別:碩士
校院名稱:亞洲大學
系所名稱:生物科技學系碩士班
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:2008
畢業學年度:96
語文別:中文
論文頁數:101
中文關鍵詞:酪胺酸酶葡萄葡萄細胞抑制作用
外文關鍵詞:tyrosinasegrapegrape cellInhibition effect
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在生物體內,當酪胺酸酶分解某些受質,如: L-3,4-dihydroxyphenylalanine (L-dopa)可導致黑色素的形成。本研究分別探討巨峰葡萄(Vitis vinifera L.kyoho)與紅地球葡萄(Vitis vinifera L. red globe)之葡萄皮、葡萄籽,以及葡萄癒傷組織細胞、紅地球葡萄籽細胞之乙醇萃取物對酪胺酸酶分解 L-dopa 活性的影響,以評估葡萄與葡萄細胞做為皮膚保養品的可行性。本實驗中酪胺酸酶活性以分解 L-dopa 在分光光度計 475 nm 下所偵測 dopachrome 的生成量為指標。實驗數據指出,葡萄籽與葡萄皮都具有抑制酪胺酸酶分解 L-dopa 的作用。巨峰葡萄籽與葡萄皮的乙醇萃取物對酪胺酸酶的抑制作用並無明顯的差異;但紅地球葡萄籽乙醇萃取物的抑制作用則遠高於其葡萄皮。故就二種葡萄整體而言,可知巨峰葡萄的乙醇萃取物之抑制作用較紅地球葡萄為佳。此外,二種葡萄細胞的乙醇萃取物僅有些微抑制酪胺酸酶活性的效果,其抑制率為 8.96 ~ 30.61 %。在酵素動力學上,各種抑制物皆對酪胺酸酶的抑制形態皆現混合型抑制作用。
Abstract
In living organisms, melanin can be formed by enzymatic reaction of tyrosinase with certain substrates such as L-3,4-dihydroxyphenylalanine (L-dopa). The objective of this study was to determine the inhibitory effects of seed and peel ethanol extracts from kyoho grape and red globe grape on tyrosinase activity. Ethanol extracts of grape cells derived from Vitis vinifera L.Gamayreaux callus and red globe grape seed were also tested to evaluate the potential of using grape as the source for skin care. The activity of tyrosinase was defined as the reactive rate to hydrolyze L-dopa into dopachrome, which can be detected at OD475. The results indicated that both the extracts from grape seed and peel showed inhibitory effect on the activity of mushroom tyrosinase. The inhibitory influence was not significant between the extracts from kyoho grape seed and peel; however, the influence was significantly higher in grape seed extract than grape peel extract in red globe grape. Therefore, ethanol extract from kyoho grape was considered to have higher inhibitory effect toward mushroom tyrosinase than ethanol extract from red globe grape. Ethanol extracts from the grape cells showed slight influence on mushroom tyrosinase activity, the inhibition rate was only 9-31%. Analysis of enzyme kinetics revealed that all the extracts showed mixed-type inhibition toward mushroom tyrosinase activity.
目錄
目錄-I
表次-IV
圖次-V
中文摘要-1
英文摘要-2
壹、前言-4
貳、文獻回顧-6
一、黑色素簡介-6
(一)、黑色素之生成-7
(二)、黑色素生成(Melanogensis)之機轉-9
(三)、黑色素之功用-13
(四)、美白機制-14
二、葡萄簡介-16
(一)、葡萄在植物學上的分類位置-16
(二)、葡萄效用-18
(三)、葡萄中已知化學成分-18
三、酵素之抑制物與其反應型態-28
(一)、競爭型抑制作用-28
(二)、非競爭型抑制作用-29
(三)、反競爭型抑制作用-30
(四)、混合型抑制作用-31
參、材料與方法-32
一、材料-32
(一)、藥品與廠商-32
(二)、實驗使用樣品-32
二、儀器-34
三、實驗方法-35
(一)、緩衝溶液配製-35
(二)、受質配製-35
(三)、酵素配製-36
(四)、抑制物的製備-36
(五)、實驗流程-37
(六)、圖表的製作-37
肆、結果與討論-40
一、各種抑制物對酪胺酸酶之 time scan 試驗-40
二、各種抑制物對酪胺酸酶之 Michaelis - Menten Kinetics 作圖-41
三、各種抑制物對酪胺酸酶之 Lineweaver - Burk plot 作圖-43
四、各種抑制物對酪胺酸酶之 Dixon 作圖-43
五、各種抑制物對酪胺酸酶活性抑制 50 % 之 IC50 試驗-44
六、各種抑制物對酪胺酸酶活性抑制之測定-45
七、各種抑制物對酪胺酸酶活性動力參數(Km)之影響-46
八、各種抑制物對酪胺酸酶活性動力參數(Vmax)之影響-47
伍、結論-49
陸、參考文獻-83
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