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標題: 聚乙二醇化對酵素活性影響之探討
Effects of PEGylation on enzyme activity
作者: 謝元榜
Hsieh, Yuan-Pang
關鍵字: PEGylation;聚乙二醇;intertwinement;amphiphilic polymers;纏繞;雙性分子
出版社: 化學工程學系所
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在此研究中,我們利用4-nitrophenyl chloroformate活化mPEG,使其接附至酵素上,經由實驗數據之分析來觀測其對酵素活性及穩定性之影響。
然本實驗選用之酵素:L-N-carbamoylase(L-Ncar)及Horseradish Peroxidase(HRP),二者經聚乙二醇化(pegylation)後,在活性上卻有相反之效果。

In this thesis, the activity and stability of PEGylated enzyme were studied. The activated mPEG (mPEG*) prepared by 4-nitrophenyl chloroformate was attached to the enzyme, and the activity of PEGylated enzymes, L-N-carbamoylase (L-Ncar) and Horseradish Peroxidase (HRP), were compared. The opposite effect of activity between two enzymes indicated the different conjugated chemistry.
The PEGylated L-Ncar shows lower activity compared to native one, and the suspicious root cause could be the long chain of mPEG causing the intertwinement with the enzyme. The long chain would cause the transformation of enzyme conformation or shield the active sites of enzyme.
On the other hand, PEGylated HRP shows higher activity with activated mPEG. This opposite result could be attributed to the hydration of mPEG. Because mPEG is an amphiphilic polymer, the effect of hydration would cause water molecules attracting to itself in the solution and, at the same time, exclude substrates from itself. It might become aqueous two-phase system in microenvironment, and it could lead to higher activity. In addition, we also found the PEGylated HRP could enhance the thermal stability and maintain high activity even in low concentration for a long term. This result implied that PEGylated HRP have more rigid conformation to resist tranformation from experimental environment.
其他識別: U0005-1708201114210000
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