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標題: 饋料式稀釋法進行乙內醯尿酶復性之研究
Refolding of D-hydantoinase with fed-batch dilution process
作者: 陳禹誠
chen, yu-cheng
關鍵字: D-hydantoinase;乙內醯尿酶;refolding
出版社: 化學工程學系所
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在利用大腸桿菌生產重組乙內醯尿酶時,其胞內會形成大量不具活性的包涵體(inclusion bodies),由於在包涵體內含有高純度的蛋白,若能有效的恢復其活性,則應用於工業上具有相當大的經濟價值。故欲發展出一個最適的復性程序,將包涵體回復成具有活性的蛋白質。實驗中先以pH 12.5, 50mM Tris-bufferru將包涵體溶解,再藉由稀釋法使其復性,並探討直接稀釋法與餽料稀釋法對乙內醯尿酶復性的影響,另外探討添加劑是否對可溶蛋白回收率有所幫助。依據實驗結果,在低流速時復性效果較佳,但添加劑對於可溶蛋白回收率並無明顯的效果。

Recombinant D-hydantoinase was over-expressed in Escherichia coli in the form of inclusion bodies, which contained large amount of the targeted proteins are valuable for industrial application if these protein could be reforded. An optimal refolding procedure has been developed to reford active proteins from inclusion bodies. The inclusion bodies were solubilized in 50mM Tris-buffer at pH 12.5 and then were diluted to refold the denatured proteins. Refolding of D-hydantoinase by direct and fed-batch operations was investigated . Furthermore, the influence of the additives, CTAB and MnCl2, for soluble protein recovery was also studied. The results of the experiments indicate that refolding process with low velocity in the fed-batch operation resulted in higher recovery of soluble protein and it is not recommended to add additives for refolding procedure.
其他識別: U0005-1708200621225700
Appears in Collections:化學工程學系所

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