Enzymatic Engineering of Live Bacterial Cell Surfaces Using Butelase 1
Enzymatic Engineering of Live Bacterial Cell Surfaces Using Butelase 1
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DOI：
10.1002/anie.201703317
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发表时间：
2017-06-26
影响因子：
16.6
通讯作者：
Liu, Chuan-Fa
中科院分区：
文献类型：
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作者：
Bi, Xiaobao;Yin, Juan;Liu, Chuan-Fa
丁酶介导的连接 (BML) 可用于用不同的货物分子修饰活细菌细胞表面。表面展示的丁酶识别基序 NHV 首先被引入到大肠杆菌细胞上锚定蛋白 OmpA 的 C 末端。然后，将其作为 BML 的手柄，使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。细胞表面连接反应是在低浓度的丁酶和标记底物下实现的。此外，荧光素标记的细菌细胞被用来显示与培养的 HeLa 细胞和活体转基因斑马鱼中巨噬细胞的相互作用，捕捉后者强大的吞噬作用。这些结果共同凸显了丁酶 1 在活细菌细胞表面工程新应用中的有用性。
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.