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Literature summary for 1.14.14.86 extracted from

  • Su, P.; Guan, H.; Zhang, Y.; Wang, X.; Gao, L.; Zhao, Y.; Hu, T.; Zhou, J.; Ma, B.; Tu, L.; Tong, Y.; Huang, L.; Gao, W.
    Probing the single key amino acid responsible for the novel catalytic function of ent-kaurene oxidase supported by NADPH-cytochrome P450 reductases in Tripterygium wilfordii (2017), Front. Plant Sci., 8, 1756 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.14.86

Cloned(Commentary)

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

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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
16alpha-hydroxy-ent-kaurane + [reduced NADPH-hemoprotein reductase] + O2 Tripterygium wilfordii
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 16alpha-hydroxy-ent-kauranoate + [oxidized NADPH-hemoprotein reductase] + H2O
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 ?
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
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 ?
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 ?
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 ?

Organism

Organism UniProt Comment Textmining
Tripterygium wilfordii
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-
-

Source Tissue

Source Tissue Comment Organism Textmining
cell suspension culture
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 Tripterygium wilfordii
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flower
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 Tripterygium wilfordii
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leaf
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 Tripterygium wilfordii
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additional information transcriptome mining Tripterygium wilfordii
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root
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 Tripterygium wilfordii
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stem
-
 Tripterygium wilfordii
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
16alpha-hydroxy-ent-kaurane + [reduced NADPH-hemoprotein reductase] + O2
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 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 ?
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 ?

Synonyms

Synonyms Comment Organism
TwKO
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 Tripterygium wilfordii

Cofactor

Cofactor Comment Organism Structure
cytochrome P450
-
 Tripterygium wilfordii
NADPH
-
 Tripterygium wilfordii

General Information

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