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Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli

Author:
Si, Tong, Tian, Qiqi, Min, Yuhao, Zhang, Linzixuan, Sweedler, Jonathan V., van der Donk, Wilfred A., Zhao, Huimin
Source:
Journal of the American Chemical Society 2018 v.140 no.38 pp. 11884-11888
ISSN:
1520-5126
Subject:
Escherichia coli, autolysis, bacteriocins, cytotoxicity, genetically modified organisms, hosts, peptides, proteinases, rapid methods, screening, signal peptide, synthetic biology
Abstract:
Most native producers of ribosomally synthesized and post-translationally modified peptides (RiPPs) utilize N-terminal leader peptides to avoid potential cytotoxicity of mature products to the hosts. Unfortunately, the native machinery of leader peptide removal is often difficult to reconstitute in heterologous hosts. Here we devised a general method to produce bioactive lanthipeptides, a major class of RiPP molecules, in Escherichia coli colonies using synthetic biology principles, where leader peptide removal is programmed temporally by protease compartmentalization and inducible cell autolysis. We demonstrated the method for producing two lantibiotics, haloduracin and lacticin 481, and performed analog screening for haloduracin. This method enables facile, high throughput discovery, characterization, and engineering of RiPPs.
Agid:
6146957