Preparative Production of Functionalized (N- and O-Heterocyclic) Polycyclic Aromatic Hydrocarbons by Human Cytochrome P450 Enzyme 3A4 in a Bioreactor

02/02/2022
 

Congratulations to Matic Srdič on his recent publication!

The integration of biocatalysis into chemical synthesis is an emerging trend that is finding increasing application in a variety of industrial processes. In Matic’s new publication, the substrate scope of the human P450 monooxygenase CYP3A4 was tested towards multiple (O/N)-polycyclic aromatic hydrocarbons (PAHs), and a bioconversion experiment was conducted in a 10 L bioreactor. This showed the synthetic potential of CYP3A4 for preparative-scale production of PAH metabolites. PAHs are dangerous environmental pollutants on one hand, and on the other hand important structural motifs in a variety of useful compounds. Functionalization of these compounds can be achieved via late-stage C-H activation by cytochrome P450 enzymes in an environmentally friendly way. The enzyme was expressed in the yeast Komagataella phaffii (Pichia pastoris) and utilized as a whole-cell biocatalyst. Several PAHs, hydroxylated PAHs, and N-PAHs were accepted as substrates, whereas none of the O-PAHs were converted. Scale-up was performed using fluorene as the model substrate in a 10 L jacketed steel bioreaction with 6 L of reaction broth volume using a common basal salts medium. The final yield was ~60% for the combined products, with ~290 mg of products formed from around ~500 mg of substrate, showing that CYP3A4 can be an excellent biocatalyst for scaled-up production of authentic human metabolites or other valuable pharmaceutical compounds.

This work has received funding from European Union’s Horizon 2020 research and innovation programme, OXYtrain MSCA-ITN, under grant agreement No 722390.

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Srdič, M., Fessner, N., Yildiz, D., Glieder, A., Spiertz, M., and Schwaneberg, U. (2022), Preparative Production of Functionalized (N- and O-Heterocyclic) Polycyclic Aromatic Hydrocarbons by Human Cytochrome P450 3A4 in a Bioreactor. Biomol., Published manuscript. https://doi.org/10.3390/biom12020153

  Overview of CYP3A4 biocatalysis scale-up strategy. Copyright: © Bio VI Overview of CYP3A4 biocatalysis scale-up strategy.