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研究生:李榮泰
研究生(外文):Jungt-Tai Li
論文名稱:有氧狀態下稻米芽鞘中胰蛋白�“磻蹌砟妞膍s
論文名稱(外文):A study of Oxygen-Dependent Protease Inhibitor from Rice Coleoptiles
指導教授:黃福永王小萍
指導教授(外文):Fu-Yung HuangShao-Pin Wang
學位類別:碩士
校院名稱:國立成功大學
系所名稱:化學系碩博士班
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:65
中文關鍵詞:芽鞘
外文關鍵詞:coleoptiles
相關次數:
  • 被引用被引用:0
  • 點閱點閱:132
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  • 下載下載:26
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胚芽米芽鞘長至3~4 公分,在通空氣時間增加的條件下,芽鞘中的
28KDa 蛋白質濃度會逐漸提高。我們利用三種方法:1.一維電泳分析後
做西方點墨法,2.利用戊二醛將Trypsin 與28KDa 蛋白質進行交聯作用
後作西方點墨法,3.經二維電泳分析後再經LC/MS 進行蛋白質序列的比
對。由上述的方法,證明28KDa 蛋白質與16KDa 一樣為Bowman-Birk 型
的胰蛋白�“磻蹌砥C
在對胚芽米分別進行缺氧與通氧不同條件下,取米粒,芽鞘、浸泡
水等各成分與DPPH(自由基的一種)進行實驗。以樣品清除率作為抗氧
化性的指標。發現在缺氧條件下的米粒,在濃度為5mg / m1 反應90 分
鐘下,具有高達73%清除率。
Plumule rice coleoptiles with the length between 3 cm and 4 cm obtained
under hypoxia condition was re-incubated under aerobic condition by purging
air. It was found that there was a 28KDa protein, which concentration
increased with the increase of air purged time, in additional to the 16KDa
protein, which already exists in coleoptiles. Three methods, western blotting
analysis, cross-linking of the 28KDa protein with the bovine trypsin using
glutaraldehyde followed by Western blotting detection, and 2-D gel
electrophoresis analysis followed by LC/MSMS analysis and N-terminal
amino acid analysis, were employed to characterize this 28 kDa protein. The
results indicated that this 28 kDa protein is a protease inhibitor and it was part
of the protein of RBBI3-1 starting from the 37th amino acid. The extracts of
the coleoptiles, the plumule rice grains after incubation under aerobic and
hypoxia conditions, and the incubating medium used in both incubation
conditions were used to assay the antioxidant activity against DPPH. It was
found that plumule rice grains after incubation in the hypoxia condition
showed 73 % activity in the concentration of 5mg / m1.
中文摘要………………………………………………………………Ⅰ
英文摘要………………………………………………………………Ⅱ
致謝……………………………………………………………………Ⅲ
表目錄…………………………………………………………………Ⅶ
圖目錄…………………………………………………………………Ⅷ
第一章 緒論
1-1 稻米基本特性……………………………………………………01
1-2 稻米的基本構造…………………………………………………03
1-3 種子發芽過程……………………………………………………04
1-4 胰蛋白�﹛K………………………………………………………06
1-5 胰蛋白�“磻蹌砥K………………………………………………10
1-6 Bowman-Birk 型抑制劑…………………………………………13
1-7 西方點墨法(Western blot)…………………………………15
1-8 蛋白質經戊二醛交聯作用………………………………………17
1-9 自由基與抗氧化…………………………………………………19
1-10 研究動機…………………………………………………………24
第二章 實驗器材與藥品
2-1 儀器………………………………………………………………25
2-2 藥品………………………………………………………………27
2-3 實驗樣品來源……………………………………………………29
第三章 實驗步驟
3-1 樣品收集…………………………………………………………30
3-2 西方點墨法………………………………………………………31
3-3 Glutaraldehyde Crosslinking測試胰蛋白��
抑制劑活性………………………………………………………33
3-4 大量樣品前處理…………………………………………………34
3-5 DEAE陰離子交換樹脂管柱………………………………………35
3-6 芽鞘蛋白質的二維電泳分析……………………………………36
3-7 膠體管柱層析……………………………………………………38
3-8 抗氧化測試………………………………………………………39
第四章 結果與討論
4-1 胚芽米通氣v.s 缺氧……………………………………………41
4-2 28KDa 的胰蛋白�“磻蹌祝〝妡P純化…………………………45
4-3 胚芽米在不同條件下抗氧化測試………………………………54
4-4 結論………………………………………………………………57
4-5 未來展望…………………………………………………………58
4-6 LC/MS 結果附表…………………………………………………59
第五章 參考文獻……………………………………………………62
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