School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.
Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore, 636921, Singapore.
Angew Chem Int Ed Engl. 2017 Jun 26;56(27):7822-7825. doi: 10.1002/anie.201703317. Epub 2017 Jun 9.
Butelase-mediated ligation (BML) can be used to modify live bacterial cell surfaces with diverse cargo molecules. Surface-displayed butelase recognition motif NHV was first introduced at the C-terminal end of the anchoring protein OmpA on E. coli cells. This then served as a handle of BML for the functionalization of E. coli cell surfaces with fluorescein and biotin tags, a tumor-associated monoglycosylated peptide, and mCherry protein. The cell-surface ligation reaction was achieved at low concentrations of butelase and the labeling substrates. Furthermore, the fluorescein-labeled bacterial cells were used to show the interactions with cultured HeLa cells and with macrophages in live transgenic zebrafish, capturing the latter's powerful phagocytic effect in action. Together these results highlight the usefulness of butelase 1 in live bacterial cell surface engineering for novel applications.
Butelase 介导的连接 (BML) 可用于用各种货物分子修饰活细菌细胞表面。将布氏菌素识别基序 NHV 首次引入大肠杆菌细胞中锚定蛋白 OmpA 的 C 末端，作为 BML 的把手，用于用荧光素和生物素标签、肿瘤相关的单糖基化肽和 mCherry 蛋白对大肠杆菌细胞表面进行功能化。在低浓度的布氏菌素和标记底物下实现了细胞表面连接反应。此外，用荧光素标记的细菌细胞与培养的 HeLa 细胞和活体转基因斑马鱼中的巨噬细胞相互作用，捕捉到后者强大的吞噬作用。这些结果共同强调了布氏菌素 1 在活细菌细胞表面工程中的有用性，可用于新的应用。
