Literature summary for 4.2.3.91 extracted from

Lopez-Gallego, F.; Agger, S.A.; Abate-Pella, D.; Distefano, M.D.; Schmidt-Dannert, C.
Sesquiterpene synthases Cop4 and Cop6 from Coprinus cinereus: catalytic promiscuity and cyclization of farnesyl pyrophosphate geometric isomers (2010), ChemBioChem, 11, 1093-1106.

Search for more literature for 4.2.3.91

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21 (DE3) cells	Coprinopsis cinerea

KM Value [mM]

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

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

Organism	UniProt	Comment	Textmining
Coprinopsis cinerea	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
Talon resin metal affinity column chromatography	Coprinopsis cinerea

Substrates and Products (Substrate)

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

Synonyms	Comment	Organism
Cop4	-	Coprinopsis cinerea

Temperature Range [°C]

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 [1/s]

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

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 Range

pH Minimum	pH Maximum	Comment	Organism
additional information	-	the catalytic fidelity of Cop4 can be drastically altered by varying reactions conditions	Coprinopsis cinerea

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Release 2023.1 (January 2023)