Cloned (Comment) | Organism |
---|---|
phylogenetic analysis, quantitative RT-PCR enzyme expression analysis, recombinant ent-kaurene oxidase expression in Saccharomyces cerevisiae strain BY-T20, coexpression with Tripterygium wilfordii CPR1-4 genes encoding four cytochrome P450 reductases, recombinant expression of enzyme mutants | Tripterygium wilfordii |
Protein Variants | Comment | Organism |
---|---|---|
L387A | site-directed mutagenesis, the mutant shows 60-70% reduced activity with 16alpha-hydroxy-ent-kaurane compared to the wild-type | Tripterygium wilfordii |
L387D | site-directed mutagenesis, the mutation causes the loss of the specific catalytic function from 16alpha-hydroxy-ent-kaurane to 16a-hydroxy-ent-kaurenoic acid | Tripterygium wilfordii |
L387G | site-directed mutagenesis, the mutant shows 60-70% reduced activity with 16alpha-hydroxy-ent-kaurane compared to the wild-type | Tripterygium wilfordii |
L387R | site-directed mutagenesis, the mutation causes the loss of the specific catalytic function from 16alpha-hydroxy-ent-kaurane to 16alpha-hydroxy-ent-kaurenoic acid | Tripterygium wilfordii |
L387S | site-directed mutagenesis, the mutant shows 60-70% reduced activity with 16alpha-hydroxy-ent-kaurane compared to the wild-type | Tripterygium wilfordii |
L387T | site-directed mutagenesis, the mutant shows 60-70% reduced activity with 16alpha-hydroxy-ent-kaurane compared to the wild-type | Tripterygium wilfordii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
16alpha-hydroxy-ent-kaurane + [reduced NADPH-hemoprotein reductase] + O2 | Tripterygium wilfordii | - |
16alpha-hydroxy-ent-kauranoate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-ene + 3 NADPH + [reduced NADPH-hemoprotein reductase] + O2 | Tripterygium wilfordii | overall reaction | ent-kaur-16-en-19-oate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
additional information | Tripterygium wilfordii | the enzyme catalyzes the three-step oxidation of the C4alpha methyl of the tetracyclic diterpene intermediate ent-kaurene to form ent-kaurenoic acid. It can also convert 16alpha-hydroxy-ent-kaurane to 16alpha-hydroxy-ent-kaurenoic acid | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Tripterygium wilfordii | - |
- |
- |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell suspension culture | - |
Tripterygium wilfordii | - |
flower | - |
Tripterygium wilfordii | - |
leaf | - |
Tripterygium wilfordii | - |
additional information | transcriptome mining | Tripterygium wilfordii | - |
root | - |
Tripterygium wilfordii | - |
stem | - |
Tripterygium wilfordii | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
16alpha-hydroxy-ent-kaurane + [reduced NADPH-hemoprotein reductase] + O2 | - |
Tripterygium wilfordii | 16alpha-hydroxy-ent-kauranoate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-ene + 3 NADPH + [reduced NADPH-hemoprotein reductase] + O2 | overall reaction | Tripterygium wilfordii | ent-kaur-16-en-19-oate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-ene + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 | overall reaction | Tripterygium wilfordii | ent-kaur-16-en-19-oate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O | - |
? | |
additional information | the enzyme catalyzes the three-step oxidation of the C4alpha methyl of the tetracyclic diterpene intermediate ent-kaurene to form ent-kaurenoic acid. It can also convert 16alpha-hydroxy-ent-kaurane to 16alpha-hydroxy-ent-kaurenoic acid | Tripterygium wilfordii | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TwKO | - |
Tripterygium wilfordii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
cytochrome P450 | - |
Tripterygium wilfordii | |
NADPH | - |
Tripterygium wilfordii |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme catalyzes the three-step oxidation of the C4alpha methyl of the tetracyclic diterpene intermediate ent-kaurene to form ent-kaurenoic acid as an early step in gibberellin (GA) biosynthesis, gibberellin phytohormone biosynthesis, pathway overview | Tripterygium wilfordii |
additional information | enzyme structur homology modeling and molecular docking, using the crystal structure of cytochrome P450 monooxygenase CYP170A1 from Streptomyces coelicolor, PDB ID 3DBG, as template. Key residue Leu387 contributes to the formation of 16alpha-hydroxy-ent-kaurenoic acid, most likely by forming hydrogen bonds with the hydroxyl group of 16alpha-hydroxy-ent-kaurane, multifunctional nature of kaurene oxidase catalysis, overview | Tripterygium wilfordii |
physiological function | the enzyme catalyzes the three-step oxidation of the C4alpha methyl of the tetracyclic diterpene intermediate ent-kaurene to form ent-kaurenoic acid as an early step in gibberellin (GA) biosynthesis. It can also convert 16alpha-hydroxy-ent-kaurane to 16alpha-hydroxy-ent-kaurenoic acid, indicating an important function of 16alpha-hydroxy-ent-kaurane in the anti-HIV principle tripterifordin biosynthetic pathway in planta | Tripterygium wilfordii |