Gee, A.R.Stone, I.Stockdale, T.P.Pukala, T.L.De Voss, J.J.Bell, S.G.2023-11-142023-11-142023Chemical Communications, 2023; 59(90):13486-134891359-73451364-548Xhttps://hdl.handle.net/2440/139892A highly sought after reaction in chemical synthesis is the activation of unactivated carbon–hydrogen bonds. We demonstrate the hydroxylation of fatty acids using an engineered thermostable archaeal cytochrome P450 enzyme. By replacing a seven amino acid section of the I-helix, the nicotinamide cofactor-dependent monooxygenase was converted into a hydrogen peroxide using peroxygenase, enabling the efficient biocatalytic oxidation of C–H bonds at room temperature to 90 °C.enThis journal is © The Royal Society of Chemistry 2023HemeCytochrome P-450 Enzyme SystemOxidation-ReductionHydroxylationBiocatalysisJournal article10.1039/d3cc04626e2023-11-14659945Gee, A.R. [0000-0001-8034-2817]Stone, I. [0000-0002-9474-4961]Pukala, T.L. [0000-0001-7391-1436]Bell, S.G. [0000-0002-7457-9727]