臺灣博碩士論文加值系統

本論文研究將澱粉修飾改質為乙醯澱粉(acetyl starch, SA)，運用在於疏水性藥物及胜肽之次微米包覆。溶於丙酮之乙醯澱粉與乳酸鏈球菌素(nisin)，再加入去離子水後即會析出包覆有Nisin之直徑為1.61μm之微顆粒其包覆率為45.5 %、吸附率為26.28 %。
介孔性分子篩SBA-15 亦可吸附大量的乳酸鏈球菌素，在pH 2條件下可吸附約2400μg /mg；pH 4吸附約2600μg /mg。而在含1M NaCl下pH2或pH 4 其乳酸鏈球菌素吸附量約為3250μg /mg。脫附試驗中在pH2溶液中脫附效果最好2600μg /mg；pH7磷酸buffer中脫附為900μg/mg，而在pH2或pH7含1M NaCl下其乳酸鏈球菌素吸脫附分別為700μg/mg及500μg /mg。在含鹽的影響下對於吸附乳酸鏈球菌素是2MNaCl>1MNaCl>0.5M NaCl。SBA-15 吸附後的乳酸鏈球菌素仍具有抑菌的活性且隨稀釋的程度而殺菌的效果也下降，並且SBA-15 吸附後的乳酸鏈球菌素在70℃加熱18小時處理後，在稀釋倍率為102，培養3小時下，抑菌效果最好，可完全抑制菌生長。

Acetyl starch , modified from corn starch was used to study the microencapsulation of hydrophobic drug and peptide.Nisin was used as the model hydrophobic peptide. Acetyl starch and Nisin can be dissolved in acetone.When water was added into the acetone solution,miroparticles with average size of 1.61μm was formed in which nisin was encapsulated.Approximately,45.5％ laded nisin was encapsulated in the starch miroparticles and about 26.3％ nisin adsorbed by the particles.
Mesoporous silica SBA-15 was also used to adsorb nisin. Approximately, 3.2mg nisin was adsorbed by 1mg of SBA-15 at pH4.0 in 1M NaCl solution. The nisin loaded SBA-15 is still possesses the antimicrobial capability. When treated at 70℃for 18h , SBA-15 could protect nisin molecules from denaturation as shown by its stable antibacterial activity

中文摘要………………………………………………………………Ⅰ
英文摘要…………….......................................Ⅱ
誌謝……………………………………………………………………Ⅲ
目錄………………..……………………………………………. ……Ⅳ
表目錄…………………………………………………………………Ⅸ
圖目錄…………………………………………………………………Ⅹ
第一章 緒論…………………………………………………………...1
1-1研究背景與目的…………………………………………………….1
1-2 研究內容簡介……………………………………………………...2
第二章 文獻回顧…………………………………..….………………..3
2.1 藥物傳輸系統…………………………………….……………… 3
2.2 澱粉……………………………………………………….……….4
2.2.1 乙醯澱粉奈米顆粒應用……………………..…………….…..6
2.3 介孔材料……………………………………………………….….7
2.3.1 SBA-15 介紹……………………….…….…………….…….9
2.3.2 SBA-15的應用……………………….………….…….…….11
V
2.4 乳酸鏈球菌素(Nisin)簡介…………………………………..….12
2.4.1 乳酸鏈球菌素(Nisin)結構………………………………..12
2.4.2 Nisin活性…………………………………………………13
2.4.3 Nisin抑菌機制…………………………………………....14
2.4.4 Nisin應用………………………………………………....16
第三章 實驗內容………………………………………………...….18
3.1實驗流程………………………………………………….…..….18
3.2 實驗材料…………………………………………………….…..21
3.2.1 菌株………………………………………………………......21
3.3實驗藥品……………………………………………….….……..21
3.4實驗設備……………………………………………………..…..22
3.5實驗培養基與試劑………………………………………..…..…23
3.5.1培養基………………………………………………….……..23
3.5.2蛋白質電泳及銀染試劑……………………………….…..…23
3.6 實驗方法…………………………………………………...……26
3.6.1乙醯澱粉包覆分析…………….……………………..…...26
3.6.1.1乙醯澱粉製備………………………………..……..….26
3.6.1.2 乙醯澱粉中乙醯取代度（DS）之測定……………..….26
VI
3.6.1.3 電子掃描式顯微鏡(SEM)樣品製備程序………………27
3.6.1.4 M17 medium 製備……………………………………...27
3.6.1.5 乳酸鏈球菌素抑菌標準檢量線建立…………...……....27
3.6.1.6 乙醯澱粉之吸附及包覆乳酸鏈球菌素之檢測………...29
3.6.1.7 不同乙醯澱粉溶液濃度、溶液濃度與水比例的包覆….29
3.6.1.8 指標菌MG1363凍菌保存………………………………30
3.6.1.9 Tricine SDS-PAGE 分析乳酸鏈球菌素………………..30
3.6.2 SBA-15 (Santa Barbara Amorphous)分析………………….33
3.6.2.1 SBA-15製備……………………………………………….33
3.6.2.2 BET比表面積分析…………………………………...…33
3.6.2.3乳酸鏈球菌素含量標準檢量線製備 ………………….....34
3.6.2.4 pH及NaCl對Nisin在 SBA-15吸附之影響………….…35
3.6.2.5乳酸鏈球菌素含NaCl 對SBA-15吸附之影響………..…35
3.6.2.6 溫度對SBA-15吸附之影響……,,,,,,,,,…………………..35
3.6.2.7 pH對Nisin在 SBA-15吸附之影響………………………36
3.6.2.8 pH及NaCl對Nisin在 SBA-15脫附之影響…………..…36
3.6.2.9 乳酸鏈球菌素抑菌活性測定………………………….....37
第四章 結果與討論………………………………………………....38
VII
4.1乙醯澱粉分析…………………………………………………….38
4.1.1乙醯澱粉中乙醯取代度（DS）之測定……………………..38
4.1.2澱粉乙醯化產率……………………………………………....39
4.1.3 乙醯澱粉的FTIR分析………………………………………40
4.1.4乙醯澱粉包覆油溶性染料試驗………………………………42
4.1.5乙醯澱粉包覆茄紅素試驗……………………………………44
4.1.6乙醯澱粉顆粒之掃描式電子顯微鏡(SEM)分析…………….45
4.7 乳酸鏈球菌素的Tricine SDS-PAGE 分析………………..….48
4.1.8乙醯澱粉之吸附及包覆乳酸鏈球菌素之檢測………………49
4.1.9不同乙醯澱粉溶液濃度、溶液濃度與水比例的包覆…..…..50
4.2 SBA-15分析……………………………………………............51
4.2.1利用穿透式電子顯微鏡(TEM)分析SBA-15…………….….51
4.2.2 SBA-15之BET測定…………………………………………52
4.2.3 pH對Nisin在 SBA-15吸附之影響……………………..….53
4.2.4 pH及NaCl對Nisin在 SBA-15吸附之影響……………....55
4.2.5 pH及NaCl對Nisin在 SBA-15脫附之影響……………….57
4.2.6乳酸鏈球菌素含NaCl 對SBA-15吸附之影響…………….59
4.2.7 Freundlich 及Langmuir 恆溫吸附線計算………………….60
4.2.8溫度對SBA-15吸附之影響…………………………………63
VIII
4.2.9 乳酸鏈球菌素在被SBA-15 吸附後的活性測試…………...65
第五章 結論與建議……………………………………….……...…67
5.1 結論……………………………………………………………...67
5.2 建議………….…………………………………………………..70
圖附錄……………………………………………………………..…71
參考文獻………………………………………………………………73
IX
表目錄
表2.1 二氧化矽介孔材料的製作特性………………………….…..8
表3.1 Tricine SDS-PAGE 膠體配置比例………………………..…31
表4.1官能基紅外線光波峰位置…………………………………….41
表4.2 電子顯微鏡下乙醯澱粉包覆不同材料之粒子觀察………...46
表4.3 乳酸鏈球菌在乙醯澱粉中包覆與吸附率…………………...50
表4.4 不同濃度乙醯澱粉、不同濃度與水比例的包覆效果………51
表4.5 SBA-15 比表面積及孔徑分佈測定………………………….53
表4.6 溶液中pH 值對SBA-15 吸附乳酸鏈球菌素量的影響…….54
表4.7 pH對Nisin在SBA-15吸附之Freundlich 恆溫吸附模式參數..62
表4.9 pH對Nisin在 SBA-15吸附之Langmuir恆溫吸附模式參數…62
表4.10 溫度對SBA-15吸附之Langmuir恆溫吸附模式參數……..64
X
圖目錄
圖2.1傳統劑型與釋放劑型的藥物濃度分佈………………..……….4
圖2.2支鏈澱粉(Amylopectin)分子結構式……………………………5
圖2.3直鏈澱粉(amylose)分子結構式…………………………...……5
圖2.4 SBA-15 合成機制……………………………………….……10
圖2.5 SBA–15形成機制示意圖……………………………….…..…11
圖2.6 乳酸鏈球菌素結構……………………………………….…..13
圖2.7 乳酸鏈球菌素之抑菌機制…………………………………...16
圖3.1乙醯澱粉包覆之實驗流程圖………………………………...19
圖3.2 SBA-15吸附/脫附實驗流程圖……………………………….20
圖3.3 96孔盤(100μL)所建立之乳酸鏈球菌素抑菌標準檢量線…29
圖3.4 Tricine SDS-PAGE 玻璃片正視圖膠之位置…………..…..32
圖3.5 乳酸鏈球菌素含量標準檢量線…………………………...…34
圖4.1 乙醯澱粉的FTIR 分析……………………………………….42
圖4.2 乙醯澱粉包覆油溶性染料試驗………………………..…….43
圖4.3 乙醯澱粉包覆油溶性染料(Blue 70)試驗……………..……..44
圖4.4 乙醯澱粉包覆茄紅素試驗………………………………..….45
圖4.5 溶液中乙醯澱粉包覆茄紅素顆粒……………………...……45
圖4.6 電子顯微鏡下乙醯澱粉形成的粒子…………………..…….47
XI
圖4.7 SEM分析乙醯澱粉包埋藍色油溶性染料形成之粒子….….47
圖4.8 SEM分析乙醯澱粉包埋乳酸鏈球菌素後形成之粒…………48
圖4.9 乳酸鏈球菌素的Tricine SDS-PAGE 分析…………………..49
圖4.10 穿透式電子顯微鏡下SBA-15 結構………………………..52
圖4.11 不同pH 情況下SBA-15 吸附乳酸鏈球菌素吸附率…...…52
圖4.12 pH及NaCl添加對SBA-15吸附Nisin的影響………………57
圖4.13 pH及NaCl添加對SBA-15脫附Nisin的影響……………....58
圖4.14 乳酸鏈球菌素含NaCl 對SBA-15 吸附效應測定……..…60
圖4.15 乳酸鏈球菌素在SBA-15恆溫吸附…………………….….61
圖4.16 乳酸鏈球菌素在不同溫度對SBA-15 吸附效應測定…....64
圖4.17 不同條件處理下乳酸鏈球菌素的抑菌能力…………….....66

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