Fernandez-Lafuente R, Armisén P, Sabuquillo P, Fernández-Lorente G, Guisán J M
Departamento de Biocatálisis, CSIC, Campus Universidad Autónoma, Madrid, Spain.
Chem Phys Lipids. 1998 Jun;93(1-2):185-97. doi: 10.1016/s0009-3084(98)00042-5.
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. continuous 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.
制备可能有助于开发有机合成工业工艺的固定化脂肪酶衍生物是一个令人兴奋的研究领域,在这个领域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兼容(例如用浓碱连续调节pH值、使用水性介质或有机溶剂等);(b) 有时,脂肪酶的一些活性/稳定性特性在固定化过程中应得到改善;(c) 由于作用机制复杂,脂肪酶在没有疏水界面的情况下活性很低。在本文中,我们将综述脂肪酶固定化的不同方法,这些方法主要与固定化衍生物在高水活性系统中进行对映选择性和区域选择性水解的进一步应用有关。特别强调脂肪酶在定制的强疏水载体表面上的选择性吸附。这种新的固定化方法基于这样一种假设,即围绕脂肪酶活性位点的大疏水区域是主要参与其在强疏水固体表面吸附的区域。因此,脂肪酶识别这些表面的方式类似于它们识别天然底物的表面,并且它们在固定化过程中会发生界面活化。这种固定化方法允许:(a) 在固定化后促进大多数脂肪酶的显著超活化。也就是说,吸附的脂肪酶在没有额外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进行高度选择性吸附。也就是说,我们可以将脂肪酶的固定化和纯化结合起来;(c) 在固定化后促进对映选择性的有趣改善;(d) 促进一种强但可逆的固定化,使我们能够在固定化脂肪酶失活后回收这些昂贵的载体。