Identification of the thiazolyl peptide GE37468 gene cluster from Streptomyces ATCC 55365 and heterologous expression in Streptomyces lividans

• DOI: 10.1073/pnas.1110435108
• Publication Date: 2011-07-26T04:02:29Z
• Authors: Travis S. Young; Christopher T. Walsh
• Keywords: Gene cluster; Heterologous expression; Heterologous; Isoleucine

Thiazolyl peptides are bacterial secondary metabolites that potently inhibit protein synthesis in Gram-positive bacteria and malarial parasites. Recently, our laboratory others reported this class of trithiazolyl pyridine-containing natural products is derived from ribosomally synthesized preproteins undergo a cascade posttranslational modifications to produce architecturally complex macrocyclic scaffolds. Here, we report the gene cluster responsible for production elongation factor Tu (EF-Tu)-targeting 29-member thiazolyl peptide GE37468 Streptomyces ATCC 55365 its heterologous expression model host lividans . harbors an unusual β -methyl- δ -hydroxy-proline residue may increase conformational rigidity macrocycle impart reduced entropic costs target binding. Isotope feeding knockout were employed engineered S. strain identify P450 monooxygenase GetJ as enzyme involved transformation isoleucine 8 through predicted tandem double hydroxylation/cyclization mechanism. Loss Ile8 oxygenative cyclization or mutation alanine via preprotein replacement resulted 4-fold 2-fold drop antibiotic activity, respectively. This genetic manipulation sets stage further examination structure activity relationships EF-Tu targeting peptides.