Regio‐ and stereoselective steroid hydroxylation by CYP109A2 from Bacillus megaterium explored by X‐ray crystallography and computational modeling

• DOI: 10.1111/febs.16906
• Publication Date: 2023-07-15T16:04:05Z
• Authors: Ilona K. Jóźwik; Elvira Bombino; Ammar Abdulmughni; Philip Hartz; Henriette J. Roze...; Hein J. Wijma; Reinhard Kappl; Dick B. Janssen; Rita Bernhardt; Andy‐Mark W. H. T...
• Keywords: Bacillus megaterium; Hydroxylation; Docking (animal)

The P450 monooxygenase CYP109A2 from Bacillus megaterium DSM319 was previously found to convert vitamin D3 (VD3) 25-hydroxyvitamin D3. Here, we show that this enzyme is also able testosterone in a highly regio- and stereoselective manner 16β-hydroxytestosterone. To reveal the structural determinants governing steroid hydroxylation reactions catalyzed by CYP109A2, two crystal structures of were solved similar closed conformations, one revealing bound active site pocket, albeit at nonproductive away heme-iron. examine whether nevertheless correspond reactive conformation docking molecular dynamics (MD) simulations performed with present site. These MD analyzed for catalytically productive relative occurrences which agreement experimentally determined stereoselectivities if predicted stability each carbon-hydrogen bond taken into account. Overall, first-time determination analysis relevant 3D will allow future small molecule ligand screening silico, as well enabling site-directed mutagenesis toward improved enzymatic properties enzyme.