Fiche publication
Date publication
juillet 2024
Journal
Microbial cell factories
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DEJAEGERE Annick
Tous les auteurs :
Puja H, Bianchetti L, Revol-Tissot J, Simon N, Shatalova A, Nommé J, Fritsch S, Stote RH, Mislin GLA, Potier N, Dejaegere A, Rigouin C
Lien Pubmed
Résumé
The engineering of non ribosomal peptide synthetases (NRPS) for new substrate specificity is a potent strategy to incorporate non-canonical amino acids into peptide sequences, thereby creating peptide diversity and broadening applications. The non-ribosomal peptide pyoverdine is the primary siderophore produced by Pseudomonas aeruginosa and holds biomedical promise in diagnosis, bio-imaging and antibiotic vectorization. We engineered the adenylation domain of PvdD, the terminal NRPS in pyoverdine biosynthesis, to accept a functionalized amino acid. Guided by molecular modeling, we rationally designed mutants of P. aeruginosa with mutations at two positions in the active site. A single amino acid change results in the successful incorporation of an azido-L-homoalanine leading to the synthesis of a new pyoverdine analog, functionalized with an azide function. We further demonstrated that copper free click chemistry is efficient on the functionalized pyoverdine and that the conjugated siderophore retains the iron chelation properties and its capacity to be recognized and transported by P. aeruginosa. The production of clickable pyoverdine holds substantial biotechnological significance, paving the way for numerous downstream applications.
Mots clés
Oligopeptides, biosynthesis, Pseudomonas aeruginosa, enzymology, Peptide Synthases, metabolism, Click Chemistry, Protein Engineering, methods, Siderophores, biosynthesis, Bacterial Proteins, metabolism, Catalytic Domain, Substrate Specificity
Référence
Microb Cell Fact. 2024 07 24;23(1):207
