Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3440
標題: PEG-HRP於有機溶劑中反應之研究
Application of PEG-modified HRP for the synthesis of polymer in organic solvent
作者: 鄭芳宜
Zheng, Fang Yi
關鍵字: horseradish peroxidase;辣根過氧化氫酶;chemical modification;monomethoxypoly(ethylene glycol);organic solvent;2-naphthol;化學修飾;甲基聚乙二醇;有機溶劑;2-奈酚
出版社: 化學工程學系
摘要: 
Pegylated horseradish peroxidase (PEG-HRP) was prepared by reacting HRP with monomethoxypoly(ethylene glycol), activated by 4-nitrophenyl chloroformate. The optimal conversion, close to 100 %, was observed in 0.1M sodium borate buffer pH9.0 with a [HRP]/[PEG] ratio of 5 x 10-3 at 25℃ for 3 hrs. The optimal activity of PEG-HRP in aqueous solution was observed at pH9.0及50℃. PEG-HRP is soluble in hydrophobic solvents such as benzene and toluene and was thus used for the polymerization of 2-naphthol in acetonitrile. The molecular weight of the polymers, exhibiting fluorescence properties, was found to increase with acetonitrile concentration.

本研究利用4-nitrophenyl chloroformate活化mPEG5000,以此活化mPEG進行辣根過氧化氫酶(Horseradish Peroxidase,HRP)化學修飾,探討最適化修飾條件及修飾化反應對HRP於水相、有機相中性質的影響,並利用修飾後HRP於有機溶劑中進行2-奈酚(2-naphthol)聚合反應。辣根過氧化氫酶最適化修飾條件為,0.1M sodium borate buffer pH9.0 ,HRP 3.5mg/ml,mPEG5000 100mg/ml,25℃反應3小時,修飾反應轉化率由SDS-PAGE分析可達100%。修飾後HRP於水相活性較未修飾HRP高,最適反應pH值與溫度分別為pH9.0及50℃,熱穩定性則沒有因修飾反應而增加。修飾後HRP可溶於Benzene、Toluene等疏水溶劑,在含50%(v/v)以上DMF或ACN與 potassium phosphate buffer混合液系統中,仍有70%以上的殘餘活性。修飾後HRP於ACN/potassium phosphate buffer混合液系統中進行2-奈酚聚合反應,產物重量平均分子量隨著ACN溶劑比例上升而增加,ACN 90%時平均分子量可達1600,螢光分析中顯示產物具有螢光反應。
URI: http://hdl.handle.net/11455/3440
Appears in Collections:化學工程學系所

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