Unusually Broad Substrate Profile of Self-Sufficient Cytochrome P450 Monooxygenase CYP116B4 from Labrenzia aggregata

A new member of the CYP116B subfamilyP450(LaMO)was discovered in Labrenzia aggregata by genomic data mining. It was successfully overexpressed in Escherichia coli, purified, and subsequently characterized spectroscopically, and its catalytic properties were assessed. Substrate profiling of the P450(LaMO) revealed that it was a versatile catalyst, exhibiting hydroxylation and epoxidation activities as well as O-dealkylation and asymmetric sulfoxidation activities. Diverse compounds, including alkylbenzenes, aromatic bicyclic molecules, and terpenoids, were shown to be hydroxylated by P450(LaMO). Such diverse catalytic activities are uncommon for the bacterial P450s, and the P450(LaMO) -mediated stereoselective hydroxylation of inactivated CH bondsubiquitous and relatively unreactive in organic moleculesis particularly unusual. The self-sufficient nature of P450(LaMO), coupled with its broad substrate range, highlights it as an ideal template for directed evolution towards various applications.