Brockelbank Jane A, Peters Verena, Rehm Bernd H A
Institute of Molecular Biosciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
Appl Environ Microbiol. 2006 Nov;72(11):7394-7. doi: 10.1128/AEM.01014-06. Epub 2006 Aug 25.
The immunoglobulin G (IgG) binding ZZ domain of protein A from Staphylococcus aureus was fused to the N terminus of the polyhydroxyalkanoate (PHA) synthase from Cupriavidus necator. The fusion protein was confirmed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and mediated formation of ZZ domain-displaying PHA granules in recombinant Escherichia coli. The IgG binding capacity of isolated granules was assessed using enzyme-linked immunosorbent assay and could be enhanced by the overproduction of the ZZ-PHA synthase. ZZ-PHA granules enabled efficient purification of IgG from human serum.
来自金黄色葡萄球菌的蛋白A的免疫球蛋白G(IgG)结合ZZ结构域与来自食酸戴尔福特菌的聚羟基脂肪酸酯(PHA)合酶的N端融合。通过基质辅助激光解吸电离飞行时间质谱法确认了融合蛋白,并介导了重组大肠杆菌中展示ZZ结构域的PHA颗粒的形成。使用酶联免疫吸附测定法评估了分离颗粒的IgG结合能力,并且可以通过过量表达ZZ-PHA合酶来增强。ZZ-PHA颗粒能够从人血清中高效纯化IgG。