Enzymatic Access to Norstatines by PLP-Dependent Decarboxylative C-C Bond Formation Involved in Leuhistin...

Norstatine (α-hydroxy-β-amino acid) is a valuable scaffold found in bioactive molecules and is particularly important as a peptidase inhibitor. Although several natural products containing norstatine scaffolds have been isolated, their biosynthetic pathways remain unknown. Here, we report two pyridoxal 5'-phosphate (PLP)-dependent enzymes, LhnD and LhnE, that catalyze norstatine formation from amino acids in the biosynthesis of leuhistin. LhnD mediates transamination of l-His to generate imidazole-5-yl pyruvate, while LhnE catalyzes a decarboxylative C-C bond formation between imidazole-5-yl pyruvate and l-Leu to complete leuhistin biosynthesis. Interestingly, isotope-labeling experiments during the LhnE reaction revealed that the catalytic cycle did not involve C_α deprotonation. This suggests the catalytic mechanism is distinct among the canonical PLP-dependent decarboxylative C-C bond forming enzymes. Genome mining of LhnE homologues also implicated the existence of previously unknown norstatine natural products. Furthermore, LhnE was successfully applied to the biocatalytic synthesis of non-natural norstatines. Together, this study provides novel insights into PLP-dependent C-C bond formation and establishes a biocatalytic route for streamlined access to norstatine pharmacophores.