1. Engineering a Highly Regioselective Fungal Peroxygenase for the Synthesis of Hydroxy Fatty Acids.
- Author
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Gomez de Santos P, González-Benjumea A, Fernandez-Garcia A, Aranda C, Wu Y, But A, Molina-Espeja P, Maté DM, Gonzalez-Perez D, Zhang W, Kiebist J, Scheibner K, Hofrichter M, Świderek K, Moliner V, Sanz-Aparicio J, Hollmann F, Gutiérrez A, and Alcalde M
- Subjects
- Oxidation-Reduction, Hydroxylation, Mixed Function Oxygenases genetics, Mixed Function Oxygenases metabolism, Fatty Acids chemistry
- 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., (© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)- Published
- 2023
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