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

Fumihiro Ishikawa; Rina Ohnishi; Chiharu Uchida; Genzoh Tanabe

doi:10.1016/j.xpro.2022.101462

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

STAR Protoc. 2022 Jun 13;3(3):101462. doi: 10.1016/j.xpro.2022.101462. eCollection 2022 Sep 16.

Authors

Fumihiro Ishikawa¹, Rina Ohnishi², Chiharu Uchida², Genzoh Tanabe³

Affiliations

¹ Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan. Electronic address: ishikawa@phar.kindai.ac.jp.
² Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan.
³ Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan. Electronic address: g-tanabe@phar.kindai.ac.jp.

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).

Keywords: Chemistry; Microbiology; Molecular/Chemical Probes; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacillus subtilis* / metabolism
Escherichia coli / genetics
Peptide Synthases* / chemistry

Substances

Peptide Synthases
non-ribosomal peptide synthase