Development of orthogonal aminoacyl-tRNA synthetase mutant for incorporating a non-canonical amino acid.
AMB Express
; 14(1): 60, 2024 May 24.
Article
in En
| MEDLINE
| ID: mdl-38782816
ABSTRACT
Genetic code expansion involves introducing non-canonical amino acids (ncAAs) with unique functional groups into proteins to broaden their applications. Orthogonal aminoacyl tRNA synthetase (aaRS), essential for genetic code expansion, facilitates the charging of ncAAs to tRNA. In this study, we developed a new aaRS mutant from Methanosaeta concilii tyrosyl-tRNA synthetase (Mc TyrRS) to incorporate para-azido-L-phenylalanine (AzF). The development involved initial site-specific mutations in Mc TyrRS, followed by random mutagenesis. The new aaRS mutant with amber suppression was isolated through fluorescence-activated cell sorting. The M. concilii aaRS mutant structure was further analyzed to interpret the effect of mutations. This research provides a novel orthogonal aaRS evolution pipeline for highly efficient ncAA incorporation that will contribute to developing novel aaRS from various organisms.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
AMB Express
Year:
2024
Document type:
Article