School of Chemistry and Manchester Interdisciplinary Biocentre, The University of Manchester, 131 Princess Street, Manchester, UKM1 7DN.
Org Biomol Chem. 2010 Feb 21;8(4):782-7. doi: 10.1039/b916773k. Epub 2009 Dec 23.
The immobilisation of proteins on to nanoparticles has a number of applications ranging from biocatalysis through to cellular delivery of biopharmaceuticals. Here we describe a phosphopantetheinyl transferase (Sfp)-catalysed method for immobilising proteins bearing a small 12-mer "ybbR" tag on to nanoparticles functionalised with coenzyme A. The Sfp-catalysed immobilisation of proteins on to nanoparticles is a highly efficient, single step reaction that proceeds under mild conditions and results in a homogeneous population of proteins that are covalently and site-specifically attached to the surface of the nanoparticles. Several enzymes of interest for biocatalysis, including an arylmalonate decarboxylase (AMDase) and a glutamate racemase (GluR), were immobilised on to nanoparticles using this approach. These enzymes retained their activity and showed high operational stability upon immobilisation.
蛋白质固定在纳米颗粒上有许多应用，从生物催化到生物制药的细胞传递。在这里，我们描述了一种磷酸泛酰巯基乙胺转移酶（Sfp）催化的方法，用于将带有小的 12 个氨基酸“ybbR”标签的蛋白质固定在辅酶 A 功能化的纳米颗粒上。Sfp 催化的蛋白质在纳米颗粒上的固定是一种高效、单步反应，在温和的条件下进行，得到的是均一的蛋白质群体，这些蛋白质通过共价键和特异性地连接到纳米颗粒的表面。几种对生物催化感兴趣的酶，包括芳基丙二酸脱羧酶（AMDase）和谷氨酸消旋酶（GluR），都使用这种方法固定在纳米颗粒上。这些酶在固定化后保留了其活性，并表现出很高的操作稳定性。
