Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis

Summary: We present an in vitro and in-cell activity-based protein profiling (ABPP) protocol for endogenous nonribosomal peptide synthetases (NRPSs). This protocol enables the fluorescence labeling and imaging of an endogenous SrfAB-NRPS with high selectivity and sensitivity in the surfactin producer Bacillus subtilis. While we optimized this protocol for use with B. subtilis, the protocol can be applied to Aneurinibacillus migulanus and Escherichia coli.For complete details on the use and execution of this protocol, please refer to Ishikawa et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.