Auto-induction medium containing glyphosate for high-level incorporation unusual aromatic amino acids into proteins

Abstract

We describe the use of an auto-induction medium containing N-(phosphono-methyl)glycine (glyphosate) as a means for high-level introduction of nonstandard aromatic amino acids into a protein. We illustrate this approach by preparing maltose binding protein (MBP) wherein all eight tryptophan residues have been replaced with 6-fluorotryptophan at an incorporation level of 99.3%. Such a high level of incorporation is important for spectroscopic investigations, in particular 19F NMR, because each species— differing amino acid sequence potentially yields a different peak pattern that complicates spectral analysis.