With the rapid development of biotechnological industry, there is an urgent need for exploiting new materials to immobilize enzymes to improve the performance of biocatalysts. In this paper, hierarchically porous magnetic microspheres (PFMMs) were prepared through solvothermal method and rapidly grafted with a novel rigid-flexible dendrimer first synthesized from monomers of trimesoyl chloride (TMC) and 1,6-hexanediamine (HDA) via interfacial polymerization process for covalent immobilization of Pseudomonas fluorescens lipase (PFL). The maximum PFL loading of the synthesized support reaches 87.5 mgprotein/gsupport, and 864% activity recovery of PFMMs-G3.0-PFL can be achieved at pH 9.0. Then, it was used to catalyze the resolution of (R,S)-1-phenylethanol with vinyl acetate. Under the optimized conditions, 50.0% conversion with 99.0% ees can be reached within 1.5 h. In addition, a conversion of 49.2% and ees of 96.9% can be retained after 10 batches of running, displaying an excellent operational stability. Importantly, a further investigation shows that the obviously improved reusability of the immobilized PFL is ascribed to the increased rigidity in comparison to fully flexible dendrimer. Thus, the newly constructed protocol for lipase immobilization exhibits a great prospect in biochemical engineering.

中文翻译：

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脂肪酶固定在新型刚性-柔性树状聚合物接枝的层级多孔磁性微球上，可有效拆分（R，S）-1-苯基乙醇。

随着生物技术产业的飞速发展，迫切需要开发新的材料来固定化酶以改善生物催化剂的性能。本文采用溶剂热法制备了多孔多孔磁性微球，并通过界面聚合工艺将其首先由均苯三甲酰氯（TMC）和1,6-己二胺（HDA）单体合成了一种新型的刚性-柔性树枝状聚合物。荧光假单胞菌脂肪酶（PFL）的共价固定化。合成载体的最大PFL负载量达到87.5 mg蛋白质/ gsupport，在pH 9.0时，可以实现864％的PFMMs-G3.0-PFL活性恢复。然后，将其用于用乙酸乙烯酯催化（R，S）-1-苯基乙醇的拆分。在优化条件下，转化率为50.0％，达到99。1.5小时内可以达到0％ee。此外，经过10批运行，可以保留49.2％的转化率和96.9％的ee，显示了出色的操作稳定性。重要的是，进一步的研究表明，与完全柔性的树枝状聚合物相比，固定式PFL明显提高的可重复使用性归因于刚性的提高。因此，新建立的脂肪酶固定方案在生化工程中具有广阔的前景。