9JQM

X-ray structure of cytochrome P450 OleT from Lacicoccus alkaliphilus in complex with icosanoic acid

Summary for 9JQM

• Entry DOI: 10.2210/pdb9jqm/pdb
• Descriptor: Fatty-acid peroxygenase, PROTOPORPHYRIN IX CONTAINING FE, icosanoic acid, ... (5 entities in total)
• Functional Keywords: decarboxylation, hydroxylation, cytochrome p450, cyp152, monooxygenase, decarboxylase, oxidoreductase
• Biological source: Lacicoccus alkaliphilus DSM 16010
• Total number of polymer chains: 2
• Total formula weight: 102638.50

Authors

Phaisan, S.,Phintha, A.,Trisrivirat, D.,Charoenpol, A.,Tanaka, H.,Kurisu, G.,Watthaisong, P.,Sucharitakul, J.,Chaiyen, P. (deposition date: 2024-09-27, release date: 2025-08-06)

Primary citation

Phaisan, S.,Phintha, A.,Trisrivirat, D.,Lawan, N.,Sucharitakul, J.,Charoenpol, A.,Watthaisong, P.,Tanaka, H.,Kurisu, G.,Chaiyen, P.
Unique structural features define the decarboxylation activity of a CYP152 fatty acid decarboxylase from Lacicoccus alkaliphilus.
J.Biol.Chem., 301:110397-110397, 2025

PubMed Abstract: Cytochrome P450 CYP152s catalyze decarboxylation of fatty acids to generate terminal alkenes, valuable compounds for various industries. Here, we identified, overexpressed, and characterized a new CYP152 enzyme from Lacicoccus alkaliphilus (OleT) and compared its biophysical and biochemical properties with the well-studied OleT from Jeotgalicoccus sp. 8456. Improved expression protocols gave the highest yields of CYP152 holoenzymes reported to date. OleT exhibits twice the catalytic turnover number of OleT when using hexadecanoic acid and HO as substrates in 10% (v/v) ethanol (EtOH). The X-ray structure of OleT in complex with icosanoic acid revealed a unique flipped heme and a substrate tunnel configuration which are different than those of other CYP152 decarboxylases. Molecular dynamics simulations revealed that in the presence of EtOH, OleT displays structural dynamics which maintain structural interactions better than those of OleT. As I178 in OleT (equivalent to L176 in OleT) shows close interactions with its substrate during simulations, I178L of OleT and L176I of OleT variants were constructed and investigated for their activities. While L176I in OleT caused a significant loss of activity, I178L of OleT had activities that were equivalent to or greater than those of the wild-type enzyme, suggesting that overall scaffold of OleT is more amenable to mutation than OleT. Stopped-flow investigations of OleT reactions indicated that EtOH increases the rate constant of Compound I formation. We also identified a new redox partner system, ferredoxin and ferredoxin reductase that can function as effective electron donors for both in vitro and in vivo systems of CYP152s.

PubMed: 40543591
DOI: 10.1016/j.jbc.2025.110397

Experimental method

X-RAY DIFFRACTION (2.44 Å)