Print Email Facebook Twitter Engineering a Highly Regioselective Fungal Peroxygenase for the Synthesis of Hydroxy Fatty Acids Title Engineering a Highly Regioselective Fungal Peroxygenase for the Synthesis of Hydroxy Fatty Acids Author Gomez de Santos, Patricia (Evoenzyme S.L., Madrid) González-Benjumea, Alejandro (CSIC - Instituto de Recursos Naturales y Agrobiologia de Sevilla (IRNAS)) Fernandez-Garcia, Angela (Institute of Catalysis, CSIC, Madrid) Aranda, Carmen (CSIC - Instituto de Recursos Naturales y Agrobiologia de Sevilla (IRNAS)) Wu, Y. (TU Delft BT/Biocatalysis) But, A. (TU Delft BT/Biocatalysis) Molina-Espeja, Patricia (Institute of Catalysis, CSIC, Madrid) Zhang, W. (TU Delft BT/Biocatalysis) Hollmann, F. (TU Delft BT/Biocatalysis) Date 2023 Abstract The hydroxylation of fatty acids is an appealing reaction in synthetic chemistry, although the lack of selective catalysts hampers its industrial implementation. In this study, we have engineered a highly regioselective fungal peroxygenase for the ω-1 hydroxylation of fatty acids with quenched stepwise over-oxidation. One single mutation near the Phe catalytic tripod narrowed the heme cavity, promoting a dramatic shift toward subterminal hydroxylation with a drop in the over-oxidation activity. While crystallographic soaking experiments and molecular dynamic simulations shed light on this unique oxidation pattern, the selective biocatalyst was produced by Pichia pastoris at 0.4 g L−1 in a fed-batch bioreactor and used in the preparative synthesis of 1.4 g of (ω-1)-hydroxytetradecanoic acid with 95 % regioselectivity and 83 % ee for the S enantiomer. Subject Hydroxy Fatty AcidsOver-OxidationProtein EngineeringRegioselectivityUnspecific Peroxygenase To reference this document use: http://resolver.tudelft.nl/uuid:30ca3257-ec6c-450a-896a-7ba54ab1c857 DOI https://doi.org/10.1002/anie.202217372 ISSN 1433-7851 Source Angewandte Chemie (International Edition), 62 (9) Part of collection Institutional Repository Document type journal article Rights © 2023 Patricia Gomez de Santos, Alejandro González-Benjumea, Angela Fernandez-Garcia, Carmen Aranda, Y. Wu, A. But, Patricia Molina-Espeja, W. Zhang, F. Hollmann, More Authors Files PDF Angew_Chem_Int_Ed_2022_Go ... sis_of.pdf 4.19 MB Close viewer /islandora/object/uuid:30ca3257-ec6c-450a-896a-7ba54ab1c857/datastream/OBJ/view