Immobilization of lipases by selective adsorption on hydrophobic supports
Immobilization of lipases by selective adsorption on hydrophobic supports
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DOI：
10.1016/s0009-3084(98)00042-5
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发表时间：
1998-06-01
影响因子：
3.4
通讯作者：
Guisán, JM
中科院分区：
文献类型：
--
作者：
Fernandez-Lafuente, R;Armisén, P;Guisán, JM
可用于开发有机合成工业过程的脂肪酶的固定化衍生物的制备是令人兴奋的研究领域，其中必须同时考虑三个主要特征：（a）固定化衍生物必须与非常不同的反应要求（例如用浓碱连续调节pH，使用水性介质或有机溶剂等）相容; (b)有时，脂肪酶的某些活性/稳定性特性应在固定化过程中得到改善;以及（c）由于复杂的作用机制，脂肪酶在不存在疏水界面的情况下活性差。本文将综述脂肪酶固定化的不同方法，主要涉及进一步使用固定化衍生物在高水活度体系中进行对映体和区域选择性水解。特别强调的是脂肪酶的选择性吸附量身定做的强疏水性支持表面。这种新的固定化过程是基于这样的假设，即脂肪酶的活性位点周围的大的疏水区域是一个主要参与它们在强疏水固体表面上的吸附。因此，脂肪酶识别这些表面类似于它们的天然底物，并且它们在固定化期间遭受界面活化。这种固定化方法允许：（a）促进大多数脂肪酶在其固定化后的显著超活化。也就是说，吸附的脂肪酶在不存在另外的疏水界面的情况下显示出非常增强的酯酶活性;（B）在非常低的离子强度下，促进脂肪酶从不纯的蛋白质提取物中的高度选择性吸附。也就是说，我们可以关联的固定化和纯化的脂肪酶;（c）促进有趣的改进固定化后的对映体选择性;和（d）促进一个强大的，但可逆的固定，使我们能够恢复这些昂贵的支持后，失活的固定化脂肪酶。(C)1998爱思唯尔科学爱尔兰有限公司保留所有权利。
The preparation of immobilized derivatives of lipases that may be useful to develop industrial processes of organic synthesis is an exciting field of research in which three main features have to be simultaneously considered: (a) immobilized derivatives have to be compatible with very different reaction requirements (e.g. continuos adjustment of pH with concentrated alkali, use of aqueous media or organic solvents, etc.); (b) Sometimes, some activity/stability properties of lipases should be improved during immobilization; and (c) because of a complex mechanism of action, lipases are poorly active in the absence of hydrophobic interfaces. In this paper, we will review different approaches for lipase immobilization mainly related to the further use of immobilized derivatives to carry out enantio and regioselective hydrolysis in high water-activity systems. Special emphasis is paid to the selective adsorption of lipases on tailor-made strongly hydrophobic support surfaces. This new immobilization procedure is based on the assumption that the large hydrophobic area that surrounds the active site of lipases is the one mainly involved in their adsorption on strongly hydrophobic solid surfaces. Thus, lipases recognize these surfaces similarly to those of their natural substrates and they suffer interfacial activation during immobilization. This immobilization method permits: (a) promote a dramatic hyper-activation of most of lipases after their immobilization. That is, adsorbed lipases show very enhanced esterase activity in the absence of additional hydrophobic interfaces; (b) promote highly selective adsorption of lipases, at very low ionic strength, from impure protein extracts. That is, we can associate immobilization and purification of lipases; (c) promote interesting improvements of enantioselectivity after immobilization; and (d) promote a strong but reversible immobilization that enables us to recover these expensive supports after inactivation of immobilized lipases. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved.