Engineering a Highly Regioselective Fungal Peroxygenase for the Synthesis of Hydroxy Fatty Acids

The hydroxylation of fatty acids is an appealing reaction in synthetic chemistry, although the lack selective catalysts hampers its industrial implementation. In this study, we have engineered a highly regioselective fungal peroxygenase for ω-1 with quenched stepwise over-oxidation. One single mutation near Phe catalytic tripod narrowed heme cavity, promoting dramatic shift toward subterminal drop over-oxidation activity. While crystallographic soaking experiments and molecular dynamic simulations shed light on unique oxidation pattern, biocatalyst was produced by Pichia pastoris at 0.4 g L−1 fed-batch bioreactor used preparative synthesis 1.4 (ω-1)-hydroxytetradecanoic acid 95 % regioselectivity 83 ee S enantiomer.