Engineering an O‐methyltransferase for the Regioselective Biosynthesis of Hesperetin Dihydrochalcone

Abstract Directed evolution of the O ‐methyltransferase ZgOMT from Zooshikella ganghwensis focusing on active site residues resulted in highly regioselective biocatalysts (regioisomeric ratios up to 99 : 1) for the preparation of the taste active hesperetin dihydrochalcone and related compounds. These newly constructed enzyme variants provide an attractive synthesis route for para ‐methylation of catechol scaffolds, which is challenging to perform with high regioselectivity utilizing wild‐type O ‐methyltransferases.