Engineered orthogonal translation systems from metagenomic libraries expand the genetic code.

ABSTRACT: Genetic code expansion with non-canonical amino acids (ncAAs) opens new opportunities for the function and design of proteins by broadening their chemical repertoire. Unfortunately, ncAA incorporation is limited both by a small collection of orthogonal aminoacyl-tRNA synthetases (aaRSs) and tRNAs and by low-throughput methods to discover them. Here, we report the discovery, characterization, and engineering of a UGA suppressing orthogonal translation system mined from metagenomic data. We developed an integrated computational and experimental pipeline to profile the orthogonality of >200 tRNAs, test >1,250 combinations of aaRS:tRNA pairs, and identify the AP1 TrpRS:tRNA^Trp _UCA as an orthogonal pair that natively encodes tryptophan at the UGA codon. We demonstrate that

SUBMITTER: Seki K 

PROVIDER: S-EPMC12636597 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature