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標題: 再生纖維素固定化金屬親和吸附薄膜於酵素右旋乙內醯尿脢純化之應用
作者: 許奇峰
關鍵字: 再生纖維素;IMAC
出版社: 化學工程學系
本研究以再生纖維素薄膜為固體載體,利用化學合成的方式接上IDA(iminodiacetic acid)螯合劑,即為固定化金屬親和薄膜(immobilized metal affinity membrane, IMAM),並利用所得之金屬親和薄膜來純化醱酵液中之右旋乙內醯尿脢(D-hydantoinase,DHTase)。文中探討DHTase吸附在薄膜上之影響因子,例如:螯合基密度、pH、NaCl的濃度等等。從實驗中我們得到最適化之操作條件,是以29.45mol Cu2+/disk-memb的膜片在0.5 M的NaCl、pH 7.5、20 mM的磷酸緩衝溶液下進行吸附,在300 mM NaCl、300 mM Imidazole、pH 8.0、20 mM的磷酸緩衝溶液狀態下進行脫附,得到最佳的右旋乙內醯尿脢(D-hydantoinase,DHTase)回收率85%,所回收酵素DHTase最佳效果為2.7mg/disk-memb,進一步做活性分析,此時具有0.0132U的活性。

The immobilized metal affinity membrane (IMAM) with modified
regeneration cellulose was employed for purification of D-hydantoinase
(DHTase).For studying DHTase adsorption capacity on IMAM, factors
such as chelator surface density, pH, NaCl concentration and elution
solutions were investigated. The optimal loading conditions were found
at 0.5M NaCl、 29.45mol Cu2+ per disk with 20mM pH 7.5 sodium
phosphate buffer .And the optimal eluted conditions were found at
0.5M NaCl、 pH8.0、 300mM imidazole、20mM phosphate buffer.
The results show that the capacity of the regeneration cellulose-based
IMAC adsorbed is 2.7mg D-hydantoinase per disk-memb.
Appears in Collections:化學工程學系所

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