葡萄糖氧化酶於強鹼性陰離子樹脂上之吸附與脫附

Adsorption and Desorption of Glucose Oxidase onto Strong-Base Anion Resins

莊家儀(Chia-I Juang)

指導教授 ： 莊瑞鑫

元智大學/工程學院/化學工程與材料科學學系/碩士(2009年)

摘要

本論文嘗試以IRA-900樹脂吸附單成分酵素溶液，實驗所使用的酵素為葡萄糖氧化酶(GOx)。而本研究共分為利用不同樹脂種類及其粒徑大小與不同緩衝液濃度、鹽類濃度和溫度，探討吸附最佳化條件、吸附熱力學，以及脫附條件之探討。
由實驗結果得知，當GOx溶於磷酸緩衝溶液中時，其pH值大於等電點，GOx帶負電。故強陰離子型交換樹脂IRA-900具有最佳之吸附效果；而在相同樹脂克數下，當樹脂粒徑越小時，其吸附量越高；隨著緩衝液濃度升高，GOx之吸附效果降低；隨著pH值、溫度升高，GOx之吸附效果隨之增高，但是活性隨著溫度上升而降低。由實驗結果得知，在使用粒徑為74μm - 250μm樹脂，吸附緩衝液為0.05 M、pH 7.8之磷酸緩衝液，並在25oC恆溫吸附下會有最佳吸附率和活性。
以Semi-reciprocal plot線性迴歸分析，確定樹脂吸附GOx符合Langmuir isothermal model，且酵素在吸附過程中的熱力學變化，均為吸熱反應。
在脫附實驗方面，使用醋酸緩衝溶液當作脫附液會造成酵素的失活，故使用磷酸緩衝溶液當作脫附液；在其條件下，增加脫附緩衝液濃度以及讓其pH值越趨近於等電點，脫附量漸增；當添加KCl濃度增高有助於酵素脫附率之提升，在1.0M KCl的脫附緩衝液濃度下，脫附率趨近100%。

關鍵字

葡萄糖氧化酶,陰離子型交換樹脂,吸附

並列摘要

In this study, the application of ion-exchange resin Amberlite IRA-900 as ion-exchange absorbent glucose oxidase (GOx). The effects of adsorption buffers concentration, pH, temperature and salt concentration on single enzyme adsorption and thermodynamics were investigated. Finally, it was discussed with desorption conditions.
For enzyme, the amount of adsorption was higher at small particle size of ion-exchange resin. The results show that the amount of enzyme adsorbed increases as the adsorption buffer concentration, temperature, pH value and salt concentration increase. In addition, the adsorption isotherms well fitted by the Langmuir isothetrmal model from the semi-reciprocal plot analysis.
For desorption condition, the amount of desorption was higher at low pH values. The results show the amount of enzyme desorbed increases as desorption buffer concentration and salt concentration increase.

並列關鍵字

Glucose Oxidase,ion-exchange resin,adsorption

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