STAR Protoc. 2026 Aug 4;7(3):104762. doi: 10.1016/j.xpro.2026.104762. Online ahead of print.
ABSTRACT
Backbone thiazole moieties (Thz) are prevalent structural features in peptidic natural products and contribute to favorable physicochemical properties. Here, we describe a protocol for in vitro chemoenzymatic synthesis of artificial Thz-containing macrocyclic peptides compatible with mRNA display workflows. We provide steps for preparing amino(thio)acid substrates and flexizyme-mediated tRNA acylation. We then detail procedures for ribosomal incorporation of thioamide linkages, spontaneous thioether macrocyclization and concomitant backbone thiazoline (Thn) formation, and enzymatic dehydrogenation of Thn to Thz to afford Thz-containing macrocycles. For complete details on the use and execution of this protocol, please refer to Saito et al.1.
PMID:42550684 | DOI:10.1016/j.xpro.2026.104762