Postgraduate Program in Biotechnology, Federal University of Alfenas, 37130-001 Alfenas, MG, Brazil.
Department of Biocatalysis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain.; Center of Excellence in Bionanoscience Research, Member of The External Scientific Advisory Board, King Abdulaziz University, Jeddah, Saudi Arabia.
Int J Biol Macromol. 2022 Aug 31;215:434-449. doi: 10.1016/j.ijbiomac.2022.06.139. Epub 2022 Jun 22.
In this review we have focused on the preparation of cross-linked enzyme aggregates (CLEAs) from lipases, as these are among the most used enzyme in bioprocesses. This immobilization method is considered very attractive due to preparation simplicity, non-use of supports and the possibility of using crude enzyme extracts. CLEAs provide lipase stabilization under extreme temperature or pH conditions or in the presence of organic solvents, in addition to preventing enzyme leaching in aqueous medium. However, it presents some problems in the preparation and limitations in their use. The problems in preparation refer mainly to the crosslinking step, and may be solved using an aminated feeder. The problems in handling have been tackled designing magnetic-CLEAs or trapping the CLEAs in particles with better mechanical properties, the substrate diffusion problems has been reduced by producing more porous-CLEAs, etc. The enzyme co-immobilization using combi-CLEAs is also a new tendency. Therefore, this review explores the CLEAs methodology aimed at lipase immobilization and its applications.
在这篇综述中，我们重点介绍了脂肪酶交联酶聚集体（CLEAs）的制备，因为这些酶是生物过程中最常用的酶之一。这种固定化方法由于制备简单、不使用载体以及可以使用粗酶提取物而被认为非常有吸引力。CLEAs 可以在极端温度或 pH 条件下或有机溶剂存在下稳定脂肪酶，同时防止酶在水介质中浸出。然而，它在制备过程中存在一些问题，并且在使用方面存在一些限制。制备过程中的问题主要涉及交联步骤，可以使用氨基供体来解决。在处理方面的问题通过设计磁性 CLEAs 或将 CLEAs 捕获在具有更好机械性能的颗粒中得到解决，通过制备更多孔的 CLEAs 等方法减少了底物扩散问题。使用组合 CLEAs 进行酶共固定化也是一个新趋势。因此，本综述探讨了旨在固定化脂肪酶及其应用的 CLEAs 方法。
