On the activity loss of hydrolases in organic solvents - I. Rapid loss of activity of a variety of enzymes and formulations in a range of organic solvents
On the activity loss of hydrolases in organic solvents - I. Rapid loss of activity of a variety of enzymes and formulations in a range of organic solvents
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DOI：
10.1016/j.molcatb.2005.06.008
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发表时间：
2005-09-01
影响因子：
--
通讯作者：
Barletta, G
中科院分区：
文献类型：
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作者：
Castillo, B;Pacheco, Y;Barletta, G
脱水酶粉末已广泛用作有机溶剂中的悬浮液来催化合成反应。要使此类应用在商业上取得成功，必须延长酶的活性。然而，最近发现许多酶的稳定性和活性在有机溶剂中受到损害。在此，我们探讨了各种水解酶（即来自梅黑毛霉和假丝酵母的脂肪酶（α-胰凝乳蛋白酶、枯草杆菌蛋白酶嘉士伯和猪肝酯酶）和各种制剂（冻干粉末、交联酶晶体、聚（乙二醇）-酶缀合物）在不同有机溶剂中的稳定性。结果显示，在暴露于有机溶剂后，所研究的所有酶和制剂的活性大致呈指数下降。观察到的失活与酶、制剂细节和溶剂无关。此外，没有发现失活程度与活性较低的制剂一样快地失去活性。与甲基-β-环糊精共冻干的枯草杆菌蛋白酶和制剂的估计半衰期 (t(1/2)) 范围为 1.8 小时到 61.6 小时。稳定的聚（乙二醇）-α-胰凝乳蛋白酶制剂。此处提供的数据表明失活可能与酶结构和动力学的变化无关。(c) 2005 Elsevier B.V. 保留所有权利。
Dehydrated enzyme powders have been used extensively as suspensions in organic solvents to catalyze synthetic reactions. Prolonged enzyme activity is necessary to make such applications commercially successful. However, it has recently become evident that the stability and thus activity of many enzymes is compromised in organic solvents. Herein we explore the stability of various hydrolases (i.e., lipases from Mucor meihei and Candida rugosa, (alpha-chymotrypsin, subtilisin Carlsberg, and pig-liver esterase) and various formulations (lyophilized powder, cross-linked enzyme crystals, poly(ethylene glycol)-enzyme conjugates) in different organic solvents. The results show a roughly exponential activity decrease for all enzymes and formulations studied after exposure to organic solvents. Inactivation was observed independent of the enzyme, formulation details, and the solvent. In addition, no relationship was found between the magnitude of inactivation and the value of initial activity. Thus, quite active formulations lost their activity as quickly as less active formulations. The estimated half-times (t(1/2)) for all enzymes and preparations ranged from 1.8 h for subtilisin C. co-lyophilized with methyl-p-cyclodextrin to 61.6 h for the most stable poly(ethylene glycol)-alpha-chymotrypsin preparation. The data here presented indicates that the inactivation is likely not related to changes in enzyme structure and dynamics. (c) 2005 Elsevier B.V. All rights reserved.