Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3) cells | Coprinopsis cinerea |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
(2E,6E)-farnesyl diphosphate | pH 8.0, 30°C | Coprinopsis cinerea | |
0.011 | - |
(2E,6E)-farnesyl diphosphate | pH not specified in the publication, at 37°C | Coprinopsis cinerea |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | replacement of Mg2+ by Mn2+ results in loss of product beta-copaene. Presence of K+ decreases the (-)-germacrene D yield and instead increases the overall yield of tricyclic sesquiterpene olefins | Coprinopsis cinerea | |
Mg2+ | - |
Coprinopsis cinerea | |
Mn2+ | replacement of Mg2+ by Mn2+ results in loss of product beta-copaene. In the presence of the Mn2+, Cop4 favors a reaction path that ends after one cyclization in (?)-germacrene D | Coprinopsis cinerea |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Coprinopsis cinerea | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Talon resin metal affinity column chromatography | Coprinopsis cinerea |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E,6E)-farnesyl diphosphate | conversion of (6E,6E)-farnesyl diphosphate proceeds via an (E,E)-germacradienyl carbocation | Coprinopsis cinerea | cubebol + diphosphate | main products at pH 8.0 are 29.3% (-)-germacrene D, 28.2% cubebol, 10.4% delta-cadinene, at pH 10.0 is 91% (-)-germacrene D, at pH 5.0 84% (-)-germacrene D plus 12% delta-cadinene | ? | |
(2E,6E)-farnesyl diphosphate + H2O | - |
Coprinopsis cinerea | cubebol + diphosphate | - |
? | |
additional information | Cop4 is a catalytically promiscuous enzyme that cyclizes (E,E)-farnesyl diphophate into multiple products, including (-)-germacrene D and cubebol (major products). Conversion of (E,E)-farnesyl diphophate by Cop4 proceeds via an (E,E)-germacradienyl carbocation. Cop4 cyclizes (2Z,6E)-farnesyl diphophate via a (6S)-beta-bisabolyl cation intermediate to (6S)-beta-bisabolene. Cop4 converts E-geranyl diphophate into cyclic and acyclic monoterpenes. Cop4 also cyclizes nerolidyl diphosphate. Under alkaline (pH 10) or acidic (pH 5.0) conditions, Cop4 becomes a very selective germacrene D synthase | Coprinopsis cinerea | ? | - |
? | |
additional information | Coprinus cinereus Cop4 additionally accepts (2Z,6E)-farnesyl diphosphate to produce 27.7% alpha-acoradiene, 10% amorpha-4,11-diene, and 13.3% cadina-4,11-diene | Coprinopsis cinerea | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Cop4 | - |
Coprinopsis cinerea |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
4 | 25 | lowering the reaction temperature from 25°C to 4°C increases the selectivity of Cop4 for (-)-germacrene D and decreases the fraction of structurally unidentified sesquiterpene olefins by half | Coprinopsis cinerea |
37 | - |
increasing the reaction temperature to 37°C decreases the fidelity of Cop4. At 37°C Cop4 generates a relative larger fraction of products beta-cubebene, sativene, delta-cadinene and beta-copaene that are derived from a cadinyl cation intermediate | Coprinopsis cinerea |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
(2E,6E)-farnesyl diphosphate | pH 8.0, 30°C | Coprinopsis cinerea | |
0.012 | - |
(2E,6E)-farnesyl diphosphate | pH not specified in the publication, at 37°C | Coprinopsis cinerea |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
under both alkaline and acidic conditions Cop4 becomes a very selective enzyme with only one major product (-)-germacrene D compared to three major compounds produced under neutral reaction conditions. At pH 10, the cyclization reaction ends with the hydride shift and deprotonation of the germacradienyl cation to yield (-)-germacrene D, 91% of total sesquiterpene products, while at pH 5.0 cyclization can proceed via a 1,6 ring closure to also yield a small amount of delta-cadinene, 12% of total sesquiterpene products, in addition to (?)-germacrene D | Coprinopsis cinerea |
5 | - |
at pH 5.0 cyclization can proceed via a 1,6 ring closure to also yield a small amount of delta-cadinene, 12% of total sesquiterpene products, in addition to (-)-germacrene D | Coprinopsis cinerea |
10 | - |
at pH 10, the cyclization reaction ends with the hydride shift and deprotonation of the germacradienyl cation to yield (-)-germacrene D, 91% of total sesquiterpene products | Coprinopsis cinerea |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the catalytic fidelity of Cop4 can be drastically altered by varying reactions conditions | Coprinopsis cinerea |