一般工業化回收基因重組大腸桿菌間質蛋白質是採用機械式方法打破細胞，然而此種操作方式會同時釋放其他胞內蛋白質，導致目標產物受到大量雜蛋白之干擾，增加後續純化步驟的複雜度。因此，滲透壓衝擊可能是回收間質蛋白質較佳之選擇。有鑑於此，本研究採取滲透壓衝擊破壞細胞外膜，自大腸桿菌間質回收肌酸酵素，同時探討在滲透壓衝擊細胞前，以鈣離子或鎂離子進行前處理對於肌酸酵素回收量、回收、與純化倍率之效應。

與傳統的滲透壓衝擊比較發現，細胞在滲透壓衝擊前以鈣離子或鎂離子進行前處理可提高肌酸酵素的回收率與純化倍率。研究結果亦顯示，細胞外膜脂多醣上之金屬結合座與鈣或鎂離子之結合並無特異性，且金屬離子與脂多醣有一吸附平衡關係。在放大實驗中發現傳統與改良之滲透壓衝擊法可處理的細胞量分別可達14 與12 g/L。
另外，本研究也比較其他回收方法對肌酸酵素回收之效應，包括以化學試劑處理的化學回收方式與添加溶菌素處理的酵素釋放方式；結果顯示以鈣離子對細胞進行前處理之滲透壓衝擊法是回收大腸桿菌間質酵素的最佳選擇。

Recovery of periplasmic enzymes from Escherichia coli on an industrial scale is normally achieved by mechanically breaking the cells. However, not only periplasmic
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As compared with the conventional osmotic shock, significant improvements on creatinase recovery and purification factor were observed when the cells pretreated with calcium or magnesium ions before the osmotic shock. The results also showed that the binding sites on the outer menbrane for metal ions were nonspecific and there was an
adsorption equilibrium relationship between lipopolysaccharides and ions. In the scale-up experiments, it was found that the maximal capability for traditional and modified osmotic shock methods were 14 and 12 g/L, respectively. In addition, this research discussed the yields of creatinase recovery by chemical permeabilization or lysozyme treatment. It indicated that the modified osmotic shock method with the pretreatment of calcium ions was a preferable choice for recovering periplasmic enzymes from E. coli cells.

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