Stereoselectivity and Substrate Specificity of the Fe(II)/α- Ketoglutarate-Dependent Oxygenase TqaL

Non-heme iron enzymes are versatile catalysts in the biosynthesis of medicinal natural products and have attracted increasing attention as practical catalytic tools in chemical synthesis due to their ability to perform chemically challenging transformations. The Fe(II)/alpha- ketoglutarate-dependent oxygenase TqaL catalyzes unusual aziridine formation from L-Val via cleavage of the unactivated C beta-H bond. However, the mechanistic details as well as the synthetic potential of TqaL-catalyzed ring closure remain unclear. Herein, we show that the TqaL-catalyzed aziridination of L-Val proceeds with an atypical, mixed stereochemical course involving both the retention and inversion of the C3(C beta) stereocenter. It is also demonstrated that TqaL accepts L-Ile and L-allo-Ile to generate the same diastereomeric pairs of aziridine products via an enzyme-controlled, stereoconvergent process. Our mutagenesis studies reveal that the reaction type (aziridination versus hydroxylation) and the stereochemical outcome are regulated by Ile343 and Phe345. Proper substitutions of Ile343 or Phe345 also make TqaL highly active toward the oxidation of alpha-amino acid substrates. This work provides mechanistic insights into the stereoselectivity and substrate specificity of the TqaL reactions.

Automated summary

Non-heme iron enzymes are versatile catalysts in the biosynthesis of medicinal natural products and have practical applications in chemical synthesis. This study provides mechanistic insights into the atypical stereochemistry of the TqaL-catalyzed aziridination reaction, which involves both retention and inversion of the C3(C beta) stereocenter. Mutagenesis studies reveal the role of specific amino acid residues in regulating the reaction type and stereochemistry.