Department of Organic Chemistry and Technology, Budapest University of Technology and Economics,1111 Budapest, Hungary.
Fermentia Microbiological Ltd., 1405 Budapest, Hungary.
Molecules. 2019 Nov 15;24(22):4146. doi: 10.3390/molecules24224146.
This article overviews the numerous immobilization methods available for various biocatalysts such as whole-cells, cell fragments, lysates or enzymes which do not require preliminary enzyme purification and introduces an advanced approach avoiding the costly and time consuming downstream processes required by immobilization of purified enzyme-based biocatalysts (such as enzyme purification by chromatographic methods and dialysis). Our approach is based on silica shell coated magnetic nanoparticles as solid carriers decorated with mixed functions having either coordinative binding ability (a metal ion complexed by a chelator anchored to the surface) or covalent bond-forming ability (an epoxide attached to the surface via a proper linker) enabling a single operation enrichment and immobilization of a recombinant phenylalanine ammonia-lyase from parsley fused to a polyhistidine affinity tag.
本文概述了各种生物催化剂（如未经初步酶纯化的完整细胞、细胞片段、细胞裂解物或酶）可用的众多固定化方法，并介绍了一种先进的方法，避免了基于纯化酶的生物催化剂固定化所需的昂贵和耗时的下游工艺（如通过色谱方法和透析进行酶纯化）。我们的方法基于涂有混合功能的二氧化硅壳包覆的磁性纳米粒子作为固体载体，这些功能具有配位结合能力（通过连接到表面的螯合剂与金属离子配位）或形成共价键的能力（通过适当的连接子将环氧化物连接到表面），能够实现单一操作从欧芹中融合了聚组氨酸亲和标签的重组苯丙氨酸氨裂解酶的富集和固定化。
