A Plug-and-Play\nT7 Expression System for Heterologous\nProduction of Lanthipeptides in Bacillus subtilis

Ribosomally synthesized lanthionine-containing peptides\n(lanthipeptides)\nhave emerged as a promising source of antimicrobials against multidrug\nresistance pathogens. An effective way to discover and engineer lanthipeptides\nis through heterologous expression of their biosynthetic gene clusters\n(BGCs) in a host of choice. Here we report a plug-and-play pathway\nrefactoring strategy for rapid evaluation of lanthipeptide BGCs in Bacillus subtilis based on the T7 expression system.\nAs a proof of concept, we used this strategy to not only observe the\nsuccessful production of a known lanthipeptide haloduracin β\nbut also discover two new human-microbiota-derived lanthipeptides\nthat previously failed to be produced in Escherichia\ncoli. The resulting B. subtilis plug-and-play T7 expression system should enable the genome mining\nof new lanthipeptides in a high-throughput manner.