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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王勝仕(Sheng-Shih Wang) | |
| dc.contributor.author | Yu-Chi Lin | en |
| dc.contributor.author | 林煜祺 | zh_TW |
| dc.date.accessioned | 2021-06-13T00:11:41Z | - |
| dc.date.available | 2007-07-31 | |
| dc.date.copyright | 2007-07-31 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-07-27 | |
| dc.identifier.citation | Batas, B. and J. B. Chaudhuri (2001). 'The influence of operational parameters on lysozyme refolding using size-exclusion chromatography.' Bioprocess and Biosystems Engineering 24(4): 255-259.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28546 | - |
| dc.description.abstract | 無論在工業上或學術研究上，對於蛋白質復性程序中，如何在高濃度蛋白質下，提高復性產率為主要課題。直接稀釋法為最簡便之復性方法，而藉由氧化還原對濃度的改變、添加劑的加入以及物理方法上的應用皆有效的提升蛋白質復性效果。
本研究針對復性緩衝液中還原態穀胱甘肽(reduced glutathione,GSH)濃度變化作分析，探討不同操作條件下所造成的影響，最後將所得濃度變化以動力學分析，期望得到一合適之反應方程式。同時將復性緩衝液隨時間放置，測量其對活性回收率之影響，討論氧化還原對(GSH/GSSG)濃度及比例在復性程序中的效果。我們使用高效能液相層析儀 (HPLC)將GSH濃度變化作分析，結果顯示在高溫環境下(50℃∼90℃)，GSH於反應後期均會產生副產物，而在低溫環境下(20℃∼40℃)，GSH則隨時間氧化成GSSG。利用動力學分析不添加EDTA時，GSH氧化速率為一0.5級反應；而添加EDTA則有效的延遲其氧化速率，為一1 級反應。在溶菌酶復性過程中，復性緩衝液隨時間放置，由於GSH氧化形成GSSG，復性效果亦隨時間增加。將GSSG濃度變化對活性回收率做圖以及GSSG/GSH比例對活性回收率做圖比較下可發現，GSSG濃度與活性回收率之間有正相關；而GSSG/GSH並非影響復性效果之絕對因素。且在GSSG濃度達1.2mM 以上時，有最好的活性回收率。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-13T00:11:41Z (GMT). No. of bitstreams: 1 ntu-96-R94524084-1.pdf: 3982922 bytes, checksum: 7ab919a1674ec85e39eff8cfe7f3a0af (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 中文摘要......................................................................I
英文摘要.....................................................................II 目錄.........................................................................IV 圖目錄.......................................................................VI 表目錄.......................................................................XI 第一章 緒論..................................................................1 第二章 文獻回顧..............................................................3 2-1-1 溶菌酶 (Lysozyme)介紹..................................................3 2-1-2 溶菌酶之活性測定.......................................................6 2-2 蛋白質之變性.............................................................7 2-2-1 蛋白質結構與穩定蛋白質分子之作用力.....................................7 2-2-2 各種蛋白質變性方法.....................................................8 2-2-3 各種常見的變性劑.......................................................9 2-3 蛋白質之復性............................................................11 2-3-1 移除變性劑............................................................13 2-3-2 降低變性劑濃度........................................................16 2-4 穀胱甘肽(glutathione)介紹...............................................27 2-4-1 穀胱甘肽(glutathione)簡介.............................................27 2-4-2 穀胱甘肽之合成(biosynthesis)及分解(degradation) ......................29 2-4-3 穀胱甘肽的功能........................................................31 2-4-4 穀胱甘肽測量方法......................................................33 第三章 研究目的.............................................................36 第四章 實驗裝置、藥品與步驟.................................................37 4-1 實驗裝置.................................................................37 4-2 實驗藥品.................................................................38 4-3 實驗步驟.................................................................39 4-3-1 以HPLC測試還原態榖胱甘肽(GSH)及氧化態榖胱甘肽(GSSG) ...................39 4-3-2 以HPLC分析復性緩衝液成分濃度變化.......................................39 4-3-3 溶菌酶之變性...........................................................42 4-3-4 溶菌酶之活性測定.......................................................42 4-3-5 溶菌酶之直接稀釋法復性.................................................43 第五章 結果與討論...........................................................44 5-1 以高效能液相層析儀(HPLC)分析復性劑之變化.................................44 5-1-1 復性緩衝液層析圖形.....................................................44 5-1-2 還原態榖胱甘肽(GSH)隨時間放置之變化...................................45 5-1-3 環境溫度對復性緩衝液之影響............................................46 5-1-4 還原態榖胱甘肽(GSH)隨時間放置之濃度變化................................48 5-2 不同操作條件下對復性緩衝液成分濃度變化之影響.............................54 5-2-1還原態榖胱甘肽(GSH)起始濃度及環境溫度之影響.............................54 5-2-2 金屬螯合劑(EDTA)之影響.................................................61 5-2-3氧化態榖胱甘肽(GSSG)對氧化速率之影響....................................68 5-3還原態榖胱甘肽(GSH)氧化速率之動力學分析...................................72 5-3-1 不添加EDTA復性緩衝液之動力學分析.......................................72 5-3-2 添加EDTA復性緩衝液之動力學分析.........................................76 5-4 復性緩衝液放置時間對復性效果之影響.......................................80 5-5 尿素對復性緩衝液之影響...................................................88 第六章 結論與建議............................................................91 參考文獻.....................................................................94 | |
| dc.language.iso | zh-TW | |
| dc.subject | 復性緩衝液 | zh_TW |
| dc.subject | 溶菌酶 | zh_TW |
| dc.subject | 穀胱甘肽 | zh_TW |
| dc.subject | 直接稀釋法 | zh_TW |
| dc.subject | lysozyme | en |
| dc.subject | refolding buffer | en |
| dc.subject | direct dilution | en |
| dc.subject | glutathione | en |
| dc.title | 復性緩衝液中氧化還原對之變化及其對溶菌酶復性效果之影響 | zh_TW |
| dc.title | The Change and Refolding Efficiency of Lysozyme of Redox Pair in Refolding Buffer | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 劉懷勝(Hwai-Shen Liu),侯劭毅,王昱麒 | |
| dc.subject.keyword | 直接稀釋法,復性緩衝液,穀胱甘肽,溶菌酶, | zh_TW |
| dc.subject.keyword | direct dilution,refolding buffer,glutathione,lysozyme, | en |
| dc.relation.page | 97 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-07-30 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
| 顯示於系所單位： | 化學工程學系 | |
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