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題名 | 探討金屬親和樹脂與金屬親和薄膜對盤尼西林醯胺酵素之純化效果=Investigation the Effect of Purified Penicillin G Acylase by Immobilized Metal Affinity Resin and Immobilized Metal Affinity Membrane |
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作者姓名(中文) | 陳志義; 姚呈儒; | 書刊名 | 修平學報 |
卷期 | 26 2013.03[民102.03] |
頁次 | 頁55-64 |
分類號 | 460.02 |
關鍵詞 | 固定化金屬親和樹脂; 親和吸附; 盤尼西林醯胺酵素; 固定化金屬親和薄膜; Immobilized metal affinity resin; IMAR; Affinity adsorption; Penicillin G acylase; PGA; Immobilized metal affinity membrane; IMAM; |
語文 | 中文(Chinese) |
中文摘要 | 本研究中,利用Novagen His-Bind resin 與RC 膜為固體載體,其帶有高密度螯合基IDA(iminodiacetic acid),可螯合銅離子,成為固定化金屬親和樹脂(immobilized metal affinity resin;IMAR)與固定化金屬親和膜(immobilized metal affinity membrane; IMAM),使用兩者對粗盤尼西林醯胺酵素液( crude penicillin G Acylase solution;crude PGA )進行親和吸附,再利用流動式反應器進行純化PGA。使用IMAR 純化PGA 之流程,清洗溶液為10mM phosphate buffer, 0.02M NH4Cl;而沖提溶液為 1M NH4Cl, 20mM imidazol , 10mM phosphate buffer ,得到78.3%回收率,10.92 倍純化倍率。IMAM 為實驗室自製,使用IMAM 純化PGA 之流程,沖提溶液為 1.5 M NH4Cl, 10mM phosphate buffer ,得到97.1%回收率,21.31 倍純化倍率。由上述結果比較,得知IMAM 純化效果較IMAR 佳。 |
英文摘要 | This study was compared the purification efficiency of immobilized metal affinity resin (IMAR) and immobilized metal affinity membrane (IMAM). Novagen His-the Bind resin and RC membrane were used as solid carrier, which contains high-density ligands of IDA (iminodiacetic acid) to chelate copper ions to become an IMAR and IMAM. Then, the IMAR and IMAM would be used to purify penicillin G acylase (PGA) form crude PGA solution with flow reactor by the effect of to affinity adsorption. For IMAR purification effect experiments, the washing solution is 10mM phosphate buffer, 0.02M of NH4Cl and the elution solution is 1M NH4Cl, 20mM imidazol, 10mM phosphate buffer. After washing and elution process, the activity recovery rate is up to 78.3% and purification increases 10.92-fold. The IMAM was laboratory-made and used the elution solution of 1.5 M NH4Cl, 10mM phosphate buffer to purify PGA. It was obtained 97.1% the activity recovery and 21.31-fold purification from the IMAM purification effect experiments. Consequently, the results showed the purification efficiency of IMAM is better than that of IMAR. |
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