8KIH

PhmA, a type I diterpene synthase without NST/DTE motif

Summary for 8KIH

• Entry DOI: 10.2210/pdb8kih/pdb
• Descriptor: diterpene synthase, PhmA, (2Z,6E,10E)-2-fluoro-3,7,11,15-tetramethylhexadeca-2,6,10,14-tetraen-1-yl trihydrogen diphosphate, MAGNESIUM ION, ... (4 entities in total)
• Functional Keywords: diterpene synthase, biosynthetic protein
• Biological source: Phoma
• Total number of polymer chains: 1
• Total formula weight: 38001.45

Authors

Zhang, B.,Ge, H.M.,Zhu, A.,Zhang, Y. (deposition date: 2023-08-23, release date: 2023-10-04, Last modification date: 2024-10-16)

Primary citation

Zhang, L.,Zhang, B.,Zhu, A.,Liu, S.H.,Wu, R.,Zhang, X.,Xu, Z.,Tan, R.X.,Ge, H.M.
Biosynthesis of Phomactin Platelet Activating Factor Antagonist Requires a Two-Enzyme Cascade.
Angew.Chem.Int.Ed.Engl., 62:e202312996-e202312996, 2023

PubMed Abstract: Phomactin diterpenoids possess a unique bicyclo[9.3.1]pentadecane skeleton with multiple oxidative modifications, and are good platelet-activating factor (PAF) antagonists that can inhibit PAF-induced platelet aggregation. In this study, we identified the gene cluster (phm) responsible for the biosynthesis of phomactins from a marine fungus, Phoma sp. ATCC 74077. Despite the complexity of their structures, phomactin biosynthesis only requires two enzymes: a type I diterpene cyclase PhmA and a P450 monooxygenase PhmC. PhmA was found to catalyze the formation of the phomactatriene, while PhmC sequentially catalyzes the oxidation of multiple sites, leading to the generation of structurally diverse phomactins. The rearrangement mechanism of the diterpene scaffold was investigated through isotope labeling experiments. Additionally, we obtained the crystal complex of PhmA with its substrate analogue FGGPP and elucidated the novel metal-ion-binding mode and enzymatic mechanism of PhmA through site-directed mutagenesis. This study provides the first insight into the biosynthesis of phomactins, laying the foundation for the efficient production of phomactin natural products using synthetic biology approaches.

PubMed: 37804495
DOI: 10.1002/anie.202312996

Experimental method

X-RAY DIFFRACTION (2 Å)