Cutting Long Syntheses Short: Access to Non‐Natural Tyrosine Derivatives Employing an Engineered Tyrosine Phenol Lyase

Abstract

AbstractThe chemical synthesis of 3‐substituted tyrosine derivatives requires a minimum of four steps to access optically enriched material starting from commercial precursors. Attempting to short‐cut the cumbersome chemical synthesis of 3‐substituted tyrosine derivatives, a single step biocatalytic approach was identified employing the tyrosine phenol lyase from Citrobacter freundii. The enzyme catalyses the hydrolysis of tyrosine to phenol, pyruvate and ammonium as well as the reverse reaction, thus the formation of tyrosine from phenol, pyruvate and ammonium. Since the wild‐type enzyme possessed a very narrow substrate spectrum, structure‐guided, site‐directed mutagenesis was required to change the substrate specificity of this CC bond forming enzyme. The best variant M379V transformed, for example, o‐cresol, o‐methoxyphenol and o‐chlorophenol efficiently to the corresponding tyrosine derivatives without any detectable side‐product. In contrast, all three phenol compounds were non‐substrates for the wild‐type enzyme. Employing the mutant, various L‐tyrosine derivatives (3‐Me, 3‐OMe, 3‐F, 3‐Cl) were obtained with complete conversion and excellent enantiomeric excess (>97%) in just a single ‘green’ step starting from pyruvate and commercially available phenol derivatives.