Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli
CABBI Theme: Conversion
Keywords: Genome Engineering, Genomics
Si, T., Tian, Q., Min, Y., Zhang, L., Sweedler, J.V., van der Donk, W.A., Zhao, H. Sept. 5, 2018. “Rapid Screening of Lanthipeptide Analogs via In-Colony Removal of Leader Peptides in Escherichia coli.” Journal of the American Chemical Society 2018, 140, 11884-11888. DOI: 10.1021/jacs.8b05544.
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.
Table S1: E. coli strains used in this study
Table S2: Plasmids used in this study and cloning strategies
Table S3: Primers used in this study