Artificial Splitting of a Non-Ribosomal Peptide Synthetase by Inserting Natural Docking Domains.

ABSTRACT: The interaction in multisubunit non-ribosomal peptide synthetases (NRPSs) is mediated by docking domains that ensure the correct subunit-to-subunit interaction. We introduced natural docking domains into the three-module xefoampeptide synthetase (XfpS) to create two to three artificial NRPS XfpS subunits. The enzymatic performance of the split biosynthesis was measured by absolute quantification of the products by HPLC-ESI-MS. The connecting role of the docking domains was probed by deleting integral parts of them. The peptide production data was compared to soluble protein amounts of the NRPS using SDS-PAGE. Reduced peptide synthesis was not a result of reduced soluble NRPS concentration but a consequence of the deletion of vital docking domain parts. Splitting the xefoampeptide biosynthe

SUBMITTER: Kegler C 

PROVIDER: S-EPMC7496407 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature