9M7F
Crystal structure of AsDMS D333N mutant in complex with farnesyl pyrophosphate
Summary for 9M7F
| Entry DOI | 10.2210/pdb9m7f/pdb |
| Descriptor | Haloacid dehalogenase superfamily, subfamily IA, variant 3 with third motif having DD or ED, FARNESYL DIPHOSPHATE, CHLORIDE ION, ... (7 entities in total) |
| Functional Keywords | terpene cyclase, haloacid dehalogenase, phosphatase, biosynthesis, biosynthetic protein |
| Biological source | Aquimarina spongiae |
| Total number of polymer chains | 2 |
| Total formula weight | 123578.82 |
| Authors | |
| Primary citation | Fujiyama, K.,Takagi, H.,Vo, N.N.Q.,Morita, N.,Nogawa, T.,Takahashi, S. Structural insights into a bacterial terpene cyclase fused with haloacid Dehalogenase-like phosphatase. Chem Sci, 16:15310-15319, 2025 Cited by PubMed Abstract: Terpene cyclases (TCs), consisting of various combinations of α, β, and γ domains, have been extensively studied. Recently, non-canonical enzymes comprising a TCβ domain and a haloacid dehalogenase (HAD)-like domain (referred to as HAD-TCβ) have been discovered. However, their overall structure remains unclear. In this study, we determined the co-crystal structures of drimenol synthase from (AsDMS), which catalyzes the conversion of farnesyl pyrophosphate (1) into drimenol (2). Crystallographic analyses of the enzyme bound to substrates 1 and drimenyl monophosphate (3) demonstrated that the TCβ domain catalyzes a class II cyclization reaction initiated by protonation, whereas the HAD domain catalyzes a phosphatase-like dephosphorylation reaction dependent on a divalent metal. Crystallographic and gel filtration analyses revealed that AsDMS adopts a dimeric assembly. This dimerization positioned the TCβ and HAD domains to facilitate efficient substrate transfer electrostatic substrate channeling. Furthermore, to investigate the structure-function relationship of the AsDMS TCβ domain, we used AlphaFold2 to model the structure of the fungal albicanol (4) synthase. Comparative analysis of active-site residues between AsDMS and fungal 4-synthase enabled rational protein engineering, converting AsDMS activity from 2-synthase to 4-synthase. This study provides insights into the biosynthesis of valuable drimane-type sesquiterpenes targeted mutagenesis. PubMed: 40852458DOI: 10.1039/d5sc04719f PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.60002725624 Å) |
Structure validation
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