Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3320
標題: 利用熱感應性高分子促進蛋白質復性之機制探討
作者: 陳怡君
Chen, Yi-Jun
關鍵字: 聚異丙基丙烯醯胺;protein;螢光特性;PNIPAAm;蛋白質;intrinsic fluorescence
出版社: 化學工程學系
摘要: 
In this study, we investigate mainly the mechanism of the poly(N-isopropylacrylamide)(PNIPAAm) assisted renaturation of carbonic anhydrase Π(CAB). It was assumed that the formation of a PNIPAAm-molten globule complex may result in a reduction in the self- association of the aggregation prone folding intermediates. This assumption was supported by the result of intrinsic fluorescence and fluorescence quenching experiments. When PNIPAAm binds to the folding intermediates, the fluorescence properties of the exposed tryptophanes are altered . The change in fluorescence properties are analogous to that observed with PEG. Not unlike PEG, an optimal stoichiometric ratio of polymer to CAB of 2 was observed. It was also found that the recovery yield can be improved by conducting refolding at lower temperature. It was assumed the strong hydrophobic interactions among folding intermediates were reduced at low temperature leading to enhance renaturation.

本研究主要探討水溶性高分子聚異丙基丙烯醯胺[poly(N-iso propyl acrylamide):PNIPAAm]輔助碳酸酐酶復性之機制,主要假設PNIPAAm會與蛋白質形成一複合體避免摺疊中間體的交互作用而發生集結以輔助蛋白質進行摺疊。
上述假設經螢光特性(intrinsic fluorescence)、螢光消光(fluorescence quenching)試驗下獲得支持,當PNIPAAm鍵結於蛋白質之一級中間體時造成表面結構的色胺酸螢光性質的改變,此結果亦相似PEG鍵結於蛋白質所造成之效應。另一方面PNIPAAm也被證實相似PEG鍵結於碳酸酶酐一級中間體時具有特殊的化學計量關係即PEG與碳酸酶酐莫耳濃度比約為2時具有最佳回收率。
同時本研究亦進行低溫狀態下添加PNIPAAm與PEG進行碳酸酐酶復性發現蛋白質復性回收率大量明顯增加並推測其原因為,降低溫度可以減緩一級中間體的強疏水基團產生交互作用,增加與高分子鍵結機會。
URI: http://hdl.handle.net/11455/3320
Appears in Collections:化學工程學系所

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