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2022•STAR ProtocolsOpen access

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

Fumihiro Ishikawa, Rina Ohnishi, Chiharu Uchida, Genzoh Tanabe

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Abstract

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).

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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).

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Available abstract

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).

Key concepts: Bacillus subtilis, Surfactin, Nonribosomal peptide, Peptide, Escherichia coli, Chemistry, Biology, Bacillaceae

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