biostudies-literatureWu KLNICHD NIH HHSNational Institute of Diabetes and Digestive and Kidney DiseasesNIAID NIH HHSBiological and Environmental ResearchNational Science Foundation of Sri LankaNational Institute of General Medical SciencesNational Institute on Minority Health and Health DisparitiesDivision of Intramural Research, National Institute of Allergy and Infectious DiseasesCancer Prevention and Research Institute of TexasU.S. Department of EnergyNIDDK NIH HHSNIMHD NIH HHSU.S. Department of DefenseNational Cancer InstituteNCI NIH HHSWelch FoundationNIGMS NIH HHSBrookhaven National Laboratorye4443https://www.ebi.ac.uk/biostudies/studies/S-EPMC9601876biostudies-literatureUnknown31(10)Genetic code expansion technology allows for the use of noncanonical amino acids (ncAAs) to create semisynthetic organisms for both biochemical and biomedical applications. However, exogenous feeding of chemically synthesized ncAAs at high concentrations is required to compensate for the inefficient cellular uptake and incorporation of these components into proteins, especially in the case of eukaryotic cells and multicellular organisms. To generate organisms capable of autonomously biosynthesizing an ncAA and incorporating it into proteins, we have engineered a metabolic pathway for the synthesis of O-methyltyrosine (OMeY). Specifically, we endowed organisms with a marformycins biosynthetic pathway-derived methyltransferase that efficiently converts tyrosine to OMeY in the presence of theProtein science : a publication of the Protein SocietyExpanding the eukaryotic genetic code with a biosynthesized 21st amino acid.PMC9601876R35-GM133706R01 DK124804W81XWH‐21‐1‐0789R35‐GM133706P30 GM133893DK124804R01 GM115261F31‐HD104474F31-HD104474DE‐SC0012704R01 AI165079P30GM133893C‐1970RR170014P30 GM138396ACB‐12002DE‐AC02‐06CH11357R01‐GM115261R01 CA217255R01-GM115261F31 HD104474STC1231306R21 CA255894R01‐AI165079P30GM138396R35 GM133706C‐2118AGM‐12006R01-CA217255R01‐CA217255RR170062R21-CA255894R01‐KP1605010R21‐CA255894Lee CDuan QPhillips GNXiao HChen YUribe RAMoore JAWu KLXu WMiller MDPeng ZfalseExpanding the eukaryotic genetic code with a biosynthesized 21st amino acid.Genetic code expansion technology allows for the use of noncanonical amino acids (ncAAs) to create semisynthetic organisms for both biochemical and biomedical applications. However, exogenous feeding of chemically synthesized ncAAs at high concentrations is required to compensate for the inefficient cellular uptake and incorporation of these components into proteins, especially in the case of eukaryotic cells and multicellular organisms. To generate organisms capable of autonomously biosynthesizing an ncAA and incorporating it into proteins, we have engineered a metabolic pathway for the synthesis of O-methyltyrosine (OMeY). Specifically, we endowed organisms with a marformycins biosynthetic pathway-derived methyltransferase that efficiently converts tyrosine to OMeY in the presence of the2022-01-01T00:00:00Z2022 Oct2026-05-28T02:02:37.779Z2025-04-05T15:22:50.932ZS-EPMC96018763617316610.1002/pro.4443