Immobilizing Penicillin G Acylase Using Silica-Supported Ionic Liquids: The Effects of Ionic Liquid Loadings

The supported ionic liquids (SILs) were prepared via immobilizing silane-functionalized ionic liquids (IL) onto the monodispersive silica nanoparticles, and the composite material was then applied in the immobilization of penicillin G acylase (PGA, EC 3.5.1.11), an important industrial enzyme for the production of semisynthetic antibiotics. This was a novel approach of immobilizing PGA proposed in our previous report, in which we had studied the effects of IL types on the performance of this novel support. Here, we focused on the effects of IL loading on the properties of the SILs and their performance in PGA immobilization. The effects of silica sizes and routes of preparing SILs (the solvent route and the sol-gel route) on IL loading were investigated. The results showed that (1) the IL loading could be tuned efficiently by changing the silica sizes. (2) As compared with the solvent route, the sol-gel route is more simple, time-saving, and ecofriendly in operation. What's more, it has a higher IL loading. Among the factors affecting the IL loading, the reaction time was the most feasible factor to control to tune the IL loading, as compared with the temperature and the dosage of ILs. (3) With the increase of IL loading, the surface zeta-potential of silicas changed from negative to positive, and the isoelectronic point (IEP) increased accordingly, which caused the decrease of the protein loading but the increase of apparent and specific activity. A reusability experiment showed that the sol-gel 1 sample maintained approximately 66% of the initial activity even after 10 consecutive operation cycles under the experimental conditions. This activity was 3.7 times higher than that of the sol-gel 2 sample.