Calleri E, Temporini C, Massolini G, Caccialanza G
Department of Pharmaceutical Chemistry, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.
J Pharm Biomed Anal. 2004 Apr 16;35(2):243-58. doi: 10.1016/S0731-7085(03)00587-9.
A review of Penicillin G Acylase (PGA)-based stationary phases is given, focusing on immobilisation methods, selection of immobilisation material and applications in chiral liquid chromatography. Two immobilization methods, namely "in situ" and "in batch" techniques, are described for the immobilisation of PGA on silica supports. Microparticulate and monolithic silica, both functionalized with aminopropyl- and epoxy-groups, were used in the development of the PGA immobilised enzyme reactor (IMER). The best results, in terms of PGA immobilised amount and enzyme activity, were obtained with the "in situ" immobilisation on epoxy monolithic silica. The use of PGA columns as enzyme reactors for the preparation of 6-APA and for the production of enantiomeric pure drugs in a one-step reaction in described. The review also covers the application of PGA-columns as chiral stationary phases for the separation of acidic enantiomers. An on-line chromatographic system based on the PGA-IMER combined with a switching valve to an analytical column is also described as a highly efficient tool to study the enantioselective hydrolyses properties of PGA. Finally a molecular modelling study is reported with the aim to give more insights into PGA-substrates interactions and to expand the application of these stationary phases as a chiral biocatalysts for pharmaceutical processes.
本文综述了基于青霉素G酰化酶（PGA）的固定相，重点介绍了固定化方法、固定化材料的选择及其在手性液相色谱中的应用。文中描述了两种将PGA固定在硅胶载体上的固定化方法，即“原位”和“分批”技术。用氨丙基和环氧基官能化的微粒硅胶和整体硅胶被用于开发PGA固定化酶反应器（IMER）。就PGA固定量和酶活性而言，在环氧整体硅胶上进行“原位”固定化可获得最佳结果。文中描述了使用PGA柱作为酶反应器，一步法制备6-氨基青霉烷酸（6-APA）和生产对映体纯药物。该综述还涵盖了PGA柱作为手性固定相用于分离酸性对映体的应用。一种基于PGA-IMER并结合切换阀连接到分析柱的在线色谱系统也被描述为研究PGA对映选择性水解特性的高效工具。最后报告了一项分子模拟研究，旨在更深入了解PGA与底物的相互作用，并扩大这些固定相作为手性生物催化剂在制药过程中的应用。
