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

Graphical Catalytic versatility of P450 La MO : A new redox-self-sufficient P450 of the CYP116B subfamily was discovered by data mining. A substrate spectrum study showed that it was a catalytic versatile monooxygenase, mediating oxidation reactions as diverse as hydroxylation, alkene epoxidation, O -dealkyl- ation, sulfoxidation, and even hydration. A new member of the CYP116B subfamily—P450 La MO —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 La MO 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 La MO . Such diverse catalytic activities are uncommon for the bacterial P450s, and the P450 La MO -mediated stereoselective hydroxylation of inactivated C  H bonds—ubiquitous and relatively unreactive in organic molecules—is particularly unusual. The self-sufficient nature of P450 La MO , coupled with its broad substrate range, highlights it as an ideal template for directed evolution towards various applications.