biostudies-literatureOlson NMNCI NIH HHSNIGMS NIH HHS110014https://www.ebi.ac.uk/biostudies/studies/S-EPMC10187777biostudies-literatureUnknown261-262Current experiments that rely on biosynthetic metabolic protein labeling with ¹⁹F often require fluorinated amino acids, which in the case of 2- and 3-fluorotyrosine can be expensive. However, using these amino acids has provided valuable insight into protein dynamics, structure, and function. Here, we develop a new in-cell method for fluorinated tyrosine generation from readily available substituted phenols and subsequent metabolic labeling of proteins in a single bacterial expression culture. This approach uses a dual-gene plasmid encoding for a model protein BRD4(D1) and a tyrosine phenol lyase from <i>Citrobacter freundii</i>, which catalyzes the formation of tyrosine from phenol, pyruvate, and ammonium. Our system demonstrated both enzymatic fluorotyrosine production and exJournal of fluorine chemistryDevelopment of a single culture <i>E. coli</i> expression system for the enzymatic synthesis of fluorinated tyrosine and its incorporation into proteins.PMC10187777P30 CA077598T32 GM008347R35 GM140837Smanski MJOlson NMPeterson KEPomerantz WCKJohnson JAHeinsch SCMarshall APCarlson EEfalseDevelopment of a single culture <i>E. coli</i> expression system for the enzymatic synthesis of fluorinated tyrosine and its incorporation into proteins.Current experiments that rely on biosynthetic metabolic protein labeling with ¹⁹F often require fluorinated amino acids, which in the case of 2- and 3-fluorotyrosine can be expensive. However, using these amino acids has provided valuable insight into protein dynamics, structure, and function. Here, we develop a new in-cell method for fluorinated tyrosine generation from readily available substituted phenols and subsequent metabolic labeling of proteins in a single bacterial expression culture. This approach uses a dual-gene plasmid encoding for a model protein BRD4(D1) and a tyrosine phenol lyase from <i>Citrobacter freundii</i>, which catalyzes the formation of tyrosine from phenol, pyruvate, and ammonium. Our system demonstrated both enzymatic fluorotyrosine production and ex2022-01-01T00:00:00Z2022 Sep2026-05-28T22:57:53.583Z2024-10-16T17:02:42.015ZS-EPMC101877773719760810.1016/j.jfluchem.2022.110014