Aminoacyl-tRNA Specificity of a Ligase Catalyzing Non-ribosomal Peptide Extension.

ABSTRACT: Peptide aminoacyl-transfer ribonucleic acid ligases (PEARLs) are amide-bond-forming enzymes that extend the main chain of peptides by using aminoacyl-tRNA (aa-tRNA) as a substrate. In this study, we investigated the substrate specificity of the PEARL BhaB_C^Ala from Bacillus halodurans, which utilizes Ala-tRNA^Ala. By leveraging flexizyme, a ribozyme capable of charging diverse acids onto a desired tRNA, we generated an array of aa-tRNAs in which we varied both the amino acid and the tRNA to dissect the substrate scope of BhaB_C^Ala. We demonstrate that BhaB_C^Ala catalyzes peptide extension with noncognate proteinogenic and noncanonical amino acids, hydroxy acids, and mercaptocarboxylic acids when attached to tRN

SUBMITTER: Nguyen DT 

PROVIDER: S-EPMC12550832 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature