Biocatalytic production of a monoamine oxidase B/catechol-O-methyltransferase inhibitor from piperine by engineered P450 BM3.

The single-step biotransformation of the natural compound piperine into a known dual inhibitor of monoamine oxidase B (MAO-B) and catechol-O-methyltransferase (COMT), was achieved by cytochrome P450 BM3 wild-type and the D251G/Q307H double mutant. This compound is used for research in neurodegenerative disorders, such as Parkinson's disease, and its value in the market is ∼14,000 €/g. Currently, it is produced by chemical synthesis requiring incubation of piperine with boron tribromide (BBr₃) in dichloromethane with yield of product not exceeding 55 % and using tedious and long procedure for its production and isolation. The P450 D251G/Q307H double mutant exhibited a 3-fold increase in catalytic efficiency compared to the wild-type enzyme, achieving high conversion (51.6 % of conversion in 15 minutes) under mild, environmentally friendly conditions. The yield of production was 0.01 mg of the inhibitor in 1 mL of reaction in 15 minutes at 28°C using the purified enzyme. Moreover, biological assays demonstrated that the resulting compound has a novel and stronger antioxidant and antimicrobial activities, respectively, when compared to piperine. The data further demonstrates the broader potential of engineered enzymes as versatile and sustainable tools in industrial biotechnology, offering an efficient platform for the modification of natural compounds to produce bioactive molecules.