The crystal structure of P450-TT heme-domain provides the first structural insights into the versatile class VII P450s

Abstract

The first crystal structure of a class VII P450, CYP116B46 from Tepidiphilus thermophilus, has been solved at 1.9 Å resolution. The structure reveals overall conservation of the P450-fold and a water conduit around the I-helix. Active site residues have been identified and sequence comparisons have been made with other class VII enzymes. A structure similarity search demonstrated that the P450-TT structure is similar to enzymes capable of oxy-functionalization of fatty acids, terpenes, macrolides, steroids and statins. The insight gained from solving this structure will provide a guideline for future engineering and modelling studies on this catalytically promiscuous class of enzymes.

Structural insight into the electron transfer pathway of a self-sufficient P450 monooxygenase

“…The helices displaying variable conformations mentioned above may move to open a tunnel to the protein surface when the active site becomes vacant 26 . The iron-heme coordination and cysteinepocket in the P450 domain of the full-length structure are the same as previously reported 27 ( Supplementary Fig. 4).…”

“…Differences are mainly observed in helix αB', αE, αF and αG, with helix αF split into αF' and αF" in the CYP116B46-N structure (helix numbering follows ref. 27 ) ( Supplementary Fig. 3).…”

“…This example highlights the potential of these self-sufficient P450s for biotechnological applications. For CYP116 family, two crystal structures of the heme domain have been reported and their substrate-binding modes were proposed accordingly 26,27 . The complex structures of a heme domain and ferredoxin from P450 systems that acquire electrons from separate redox partners have also been reported [28][29][30][31] , which demonstrated how the electrons are injected from [2Fe-2S] cluster to the heme (Supplementary Fig.…”

Regio‐ and Enantio‐selective Chemo‐enzymatic C−H‐Lactonization of Decanoic Acid to (S)‐δ‐Decalactone

“…We were especially interested in the excellent regioselectivity for the challenging C5 hydroxylation of decanoic acid to 5‐hydroxy decanoic acid displayed by P450‐TT because this reactivity creates precursor 2 for the valuable δ‐decalactone ( 3 ). Using the recently solved crystal structure of P450‐TT, molecular docking simulations of decanoic acid into the heme domain of P450‐TT were performed (Figure ). The energetically most favored simulation creates a reasonable binding pose for promoting substrate hydroxylation .…”

Regio‐ and Enantio‐selective Chemo‐enzymatic C−H‐Lactonization of Decanoic Acid to (S)‐δ‐Decalactone

“…We were especially interested in the excellent regioselectivity for the challenging C5 hydroxylation of decanoic acid to 5-hydroxy decanoic acid displayed by P450-TT because this reactivity creates precursor 2 for the valuable d-decalactone (3). Using the recently solved crystal structure of P450-TT, [36] molecular docking simulations of decanoic acid into the heme domain of P450-TT were performed (Figure 1). Thee nergetically most favored simulation creates ar easonable binding Scheme 1.…”