Cloned (Comment) | Organism |
---|---|
expression in Pichia pastoris | Physcomitrium patens |
gene CYP701B1, genetic structure, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzyme in Pichia pastoris microsomes | Physcomitrium patens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | among the azoles known as CYP51 inhibitors, ketoconazole, an antifungal medicine, is one of the most potent CYP51 inhibitor and forms a one-to-one complex with CYP51 as with uniconazole and CYP701B1. But ketoconazole does not bind and inhibit the purified ent-kaurene oxidase, CYP701B1; not inhibitory: ketoconazole | Physcomitrium patens | |
uniconazole | selective binding of CYP701B1 with uniconazole, is directly bound to the heme iron with its azole nitrogen | Physcomitrium patens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | enzyme kinetics | Physcomitrium patens | |
0.0091 | - |
ent-kaur-16-ene | pH 7.5, 30°C | Physcomitrium patens | |
0.0091 | - |
ent-kaur-16-ene | pH 7.5, 30°C, recombinant His-tagged enzyme | Physcomitrium patens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
microsome | - |
Physcomitrium patens | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | purified ferric CYP701B1 is a typical low-spin cytochrome P450 showing a characteristic absorption peak | Physcomitrium patens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ent-kaur-16-en-19-al + [reduced NADPH-hemoprotein reductase] + O2 | Physcomitrium patens | - |
ent-kaur-16-en-19-oate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-en-19-ol + [reduced NADPH-hemoprotein reductase] + O2 | Physcomitrium patens | - |
ent-kaur-16-en-19-al + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-ene + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 | Physcomitrium patens | overall reaction | ent-kaur-16-en-19-oate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O | - |
? | |
ent-kaur-16-ene + [reduced NADPH-hemoprotein reductase] + O2 | Physcomitrium patens | - |
ent-kaur-16-en-19-ol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Physcomitrium patens | - |
- |
- |
Physcomitrium patens | F2ZAE0 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Pichia pastoris microsomes by ultracentrifugation, nickel affinity chromatography, and dialysis | Physcomitrium patens |
recombinant protein | Physcomitrium patens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ent-kaur-16-en-19-al + [reduced NADPH-hemoprotein reductase] + O2 | - |
Physcomitrium patens | ent-kaur-16-en-19-oate + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-en-19-ol + [reduced NADPH-hemoprotein reductase] + O2 | - |
Physcomitrium patens | ent-kaur-16-en-19-al + [oxidized NADPH-hemoprotein reductase] + 2 H2O | - |
? | |
ent-kaur-16-en-19-ol + [reduced NADPH-hemoprotein reductase] + O2 | - |
Physcomitrium patens | ent-kaur-16-en-19-al + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
ent-kaur-16-ene + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 | - |
Physcomitrium patens | ent-kaur-16-en-19-oate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O | - |
? | |
ent-kaur-16-ene + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 | overall reaction | Physcomitrium patens | ent-kaur-16-en-19-oate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O | - |
? | |
ent-kaur-16-ene + 3 [reduced NADPH-hemoprotein reductase] + 3 O2 | overall reaction, substrate is isolated from leaves of Cryptomeria japonica | Physcomitrium patens | ent-kaur-16-en-19-oate + 3 [oxidized NADPH-hemoprotein reductase] + 4 H2O | - |
? | |
ent-kaur-16-ene + [reduced NADPH-hemoprotein reductase] + O2 | - |
Physcomitrium patens | ent-kaur-16-en-19-ol + [oxidized NADPH-hemoprotein reductase] + H2O | - |
? | |
additional information | enzyme catalyzes the oxidation of ent-kaurene to entkaurenoic acid through three successive monooxygenations. Te rate-limiting step of this oxidation might be the initial step that forms ent-kaurenol | Physcomitrium patens | ? | - |
? | |
additional information | GC-MS or GLC product identification | Physcomitrium patens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 62000, SDS-PAGE | Physcomitrium patens |
Synonyms | Comment | Organism |
---|---|---|
CYP701B1 | - |
Physcomitrium patens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Physcomitrium patens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Physcomitrium patens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
cytochrome P450 | - |
Physcomitrium patens | |
cytochrome P450 | typical ferric low-spin cytochrome P450, which is completely moved to high-spin state upon binding with ent-kaurene | Physcomitrium patens | |
heme | - |
Physcomitrium patens | |
NADPH | - |
Physcomitrium patens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.00003 | - |
pH 7.5, 30°C | Physcomitrium patens | uniconazole |
General Information | Comment | Organism |
---|---|---|
metabolism | oxidation of ent-kaurene to ent-kaurenoic acid is a common process in gibberellin biosynthesis by vascular plants and production of gibberellin-like compounds by nonvascular plants | Physcomitrium patens |