| IED ID | IndEnz0010001854 |
| Enzyme Type ID | esterase001854 |
| Protein Name |
Trichodiene synthase EC 4.2.3.6 Core trichothecene cluster CTC protein 5 Sesquiterpene cyclase TRI5 TS |
| Gene Name | TRI5 TOX 5 |
| Organism | Fusarium sporotrichioides |
| Taxonomic Lineage | cellular organisms Eukaryota Opisthokonta Fungi Dikarya Ascomycota saccharomyceta Pezizomycotina leotiomyceta sordariomyceta Sordariomycetes Hypocreomycetidae Hypocreales Nectriaceae Fusarium Fusarium sambucinum species complex Fusarium sporotrichioides |
| Enzyme Sequence | MENFPTEYFLNTTVRLLEYIRYRDSNYTREERIENLHYAYNKAAHHFAQPRQQQLLKVDPKRLQASLQTIVGMVVYSWAKVSKECMADLSIHYTYTLVLDDSKDDPYPTMVNYFDDLQAGREQAHPWWALVNEHFPNVLRHFGPFCSLNLIRSTLDFFEGCWIEQYNFGGFPGSHDYPQFLRRMNGLGHCVGASLWPKEQFNERSLFLEITSAIAQMENWMVWVNDLMSFYKEFDDERDQISLVKNYVVSDEISLHEALEKLTQDTLHSSKQMVAVFSDKDPQVMDTIECFMHGYVTWHLCDRRYRLSEIYEKVKEEKTEDAQKFCKFYEQAANVGAVSPSEWAYPPVAQLANVRSKDVKEVQKPFLSSIELVE |
| Enzyme Length | 374 |
| Uniprot Accession Number | P13513 |
| Absorption | |
| Active Site | |
| Activity Regulation | ACTIVITY REGULATION: Benzyl triethylammonium cation (BTAC) acts as a competitive inhibitor of trichodiene synthase reaction in the presence of pyrophosphate (PPi) (PubMed:17678871). {ECO:0000269|PubMed:17678871}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=(2E,6E)-farnesyl diphosphate = diphosphate + trichodiene; Xref=Rhea:RHEA:12052, ChEBI:CHEBI:15861, ChEBI:CHEBI:33019, ChEBI:CHEBI:175763; EC=4.2.3.6; Evidence={ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17678871, ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:7873527, ECO:0000269|PubMed:8823172}; |
| DNA Binding | |
| EC Number | 4.2.3.6 |
| Enzyme Function | FUNCTION: Trichodiene synthase; part of the core gene cluster that mediates the biosynthesis of trichothecenes, a very large family of chemically related bicyclic sesquiterpene compounds acting as mycotoxins, including T2-toxin (PubMed:11352533, PubMed:2817906). The biosynthesis of trichothecenes begins with the cyclization of farnesyl diphosphate to trichodiene and is catalyzed by the trichodiene synthase TRI5 (PubMed:3800398, PubMed:7873527, PubMed:8823172). Trichodiene undergoes a series of oxygenations catalyzed by the cytochrome P450 monooxygenase TRI4 (PubMed:7651333). TRI4 controls the addition of four oxygens at C-2, C-3, C-11, and the C-12, C-13-epoxide to form the intermediate isotrichotriol (PubMed:16917519). Isotrichotriol then undergoes a non-enzymatic isomerization and cyclization to form isotrichodermol (PubMed:2317042). During this process, the oxygen at the C-2 position becomes the pyran ring oxygen and the hydroxyl group at C-11 is lost (PubMed:2317042). More complex type A trichothecenes are built by modifying isotrichodermol through a series of paired hydroxylation and acetylation or acylation steps (PubMed:11352533). Isotrichodermol is converted to isotrichodermin by the acetyltransferase TRI101 (PubMed:10583973). TRI101 encodes a C-3 transacetylase that acts as a self-protection or resistance factor during biosynthesis and that the presence of a free C-3 hydroxyl group is a key component of Fusarium trichothecene phytotoxicity (PubMed:10583973). A second hydroxyl group is added to C-15 by the trichothecene C-15 hydroxylase TRI11, producing 15-decalonectrin, which is then acetylated by TRI3, producing calonectrin (PubMed:9435078, PubMed:8593041). A third hydroxyl group is added at C-4 by the cytochrome P450 monooxygenase TRI13, converting calonectrin to 3,15-diacetoxyspirpenol, which is subsequently acetylated by the acetyltransferase TRI7 (PubMed:12135578, PubMed:11352533). A fourth hydroxyl group is added to C-8 by the cytochrome P450 monooxygenase TRI1, followed by the addition of an isovaleryl moiety by TRI16 (PubMed:12620849, PubMed:14532047). Finally, the acetyl group is removed from the C-3 position by the trichothecene C-3 esterase TRI8 to produce T-2 toxin (PubMed:12039755). {ECO:0000269|PubMed:10583973, ECO:0000269|PubMed:11352533, ECO:0000269|PubMed:12039755, ECO:0000269|PubMed:12135578, ECO:0000269|PubMed:12620849, ECO:0000269|PubMed:14532047, ECO:0000269|PubMed:16917519, ECO:0000269|PubMed:2317042, ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:7651333, ECO:0000269|PubMed:8593041, ECO:0000269|PubMed:9435078}. |
| Temperature Dependency | |
| PH Dependency | BIOPHYSICOCHEMICAL PROPERTIES: pH dependence: Optimum pH is 6.75-7.75. {ECO:0000269|PubMed:3800398}; |
| Pathway | PATHWAY: Sesquiterpene biosynthesis; trichothecene biosynthesis. {ECO:0000269|PubMed:3800398}. |
| nucleotide Binding | |
| Features | Beta strand (1); Chain (1); Helix (20); Metal binding (7); Mutagenesis (11); Region (1); Turn (3) |
| Keywords | 3D-structure;Direct protein sequencing;Lyase;Magnesium;Metal-binding |
| Interact With | |
| Induction | INDUCTION: Expression is positively regulated by the trichothecene cluster-specific transcription activator TRI10 (PubMed:12732543). Expression is induced between 18h and 21h growth on GYEP medium (PubMed:16347944). The initial detection of trichothecenes occurs several hours after the initial detection of TRI5 (PubMed:16347944). The initiation of trichothecene biosynthesis occurs with a high concentration of glucose remaining in the culture medium (PubMed:16347944). {ECO:0000269|PubMed:12732543, ECO:0000269|PubMed:16347944}. |
| Subcellular Location | |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | X-ray crystallography (20) |
| Cross Reference PDB | 1JFA; 1JFG; 1KIY; 1KIZ; 1YJ4; 1YYQ; 1YYR; 1YYS; 1YYT; 1YYU; 2AEK; 2AEL; 2AET; 2PS4; 2PS5; 2PS6; 2PS7; 2PS8; 2Q9Y; 2Q9Z; |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 44,000 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=62.0 nM for farnesyl pyrophosphate {ECO:0000269|PubMed:3800398, ECO:0000269|PubMed:7873527}; KM=78.0 nM for Mg(2+) {ECO:0000269|PubMed:8823172}; KM=84.8 nM for Mn(2+) {ECO:0000269|PubMed:8823172}; |
| Metal Binding | METAL 100; /note="Magnesium 1"; /evidence="ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2PS7"; METAL 164; /note="Magnesium 1"; /evidence="ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2PS7"; METAL 225; /note="Magnesium 2"; /evidence="ECO:0000269|PubMed:15835903, ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718, ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR, ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET, ECO:0007744|PDB:2PS8"; METAL 229; /note="Magnesium 2"; /evidence="ECO:0000269|PubMed:15835903, ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718, ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR, ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET, ECO:0007744|PDB:2PS8"; METAL 233; /note="Magnesium 2"; /evidence="ECO:0000269|PubMed:15835903, ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718, ECO:0007744|PDB:1YYQ, ECO:0007744|PDB:1YYR, ECO:0007744|PDB:1YYU, ECO:0007744|PDB:2AET, ECO:0007744|PDB:2PS8"; METAL 239; /note="Magnesium 3"; /evidence="ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2AEK, ECO:0007744|PDB:2PS4"; METAL 241; /note="Magnesium 3; via carbonyl oxygen"; /evidence="ECO:0000269|PubMed:16171386, ECO:0000269|PubMed:17996718, ECO:0007744|PDB:2AEK, ECO:0007744|PDB:2PS4" |
| Rhea ID | RHEA:12052 |
| Cross Reference Brenda | 4.2.3.6; |