In-Situ Purification of Non-Ribosomal Peptide Synthetases Assembly Line for Structural and Biochemical Studies.

ABSTRACT: Nonribosomal peptide synthetases (NRPS) are essential for the biosynthesis of therapeutically valuable molecules, including antibiotics, immunosuppressants, and anticancer agents. The assembly-line mechanism of NRPS offers significant potential for engineering novel natural products through reprogramming. However, the challenging purification of NRPS proteins has impeded the investigation of their assembly and catalytic mechanisms. In this study, we employed homologous recombination to insert a purification tag at the C-terminus of the NRPS gene within the chromosome. This genetic modification enabled efficient purification of NRPS proteins from the tagged mutant strain using a one-step affinity chromatography approach. Additionally, we discovered that MbtH-like proteins (MLPs) form stable

SUBMITTER: Cao W 

PROVIDER: S-EPMC11855355 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature