Bacterial CYP153A monooxygenases for the synthesis of omega-hydroxylated fatty acidsElectronic supplementary information (ESI) available: Materials and methods. Detailed substrate conversions and product distributions. Gas chromatograms and mass spectra. Michaelis-Menten plots. See DOI: 10.1039/c2cc18103g

CYP153A from Marinobacter aquaeolei has been identified as a fatty acid ω-hydroxylase with a broad substrate range. Two hotspots predicted to influence substrate specificity and selectivity were exchanged. Mutant G307A is 2- to 20-fold more active towards fatty acids than the wild-type. Residue L354 is determinant for the enzyme ω-regioselectivity. A CYP153A monooxygenase has been characterized as a fatty acid ω-hydroxylase with a broad substrate range. Using a sequence-based approach, two hotspots were identified as pivotal for activity and selectivity.