Information on EC 4.2.3.95 - (-)-alpha-cuprenene synthase

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The expected taxonomic range for this enzyme is: Coprinopsis cinerea

EC NUMBER
COMMENTARY hide
4.2.3.95
-
RECOMMENDED NAME
GeneOntology No.
(-)-alpha-cuprenene synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2E,6E)-farnesyl diphosphate = (-)-alpha-cuprenene + diphosphate
show the reaction diagram
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
(-)-alpha-cuprenene hydrolase [cyclizing, (-)-alpha-cuprenene-forming]
The enzyme from the fungus Coprinopsis cinerea produces (-)-alpha-cuprenene with high selectivity.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2E,6E)-farnesyl diphosphate
(-)-alpha-cuprenene + diphosphate
show the reaction diagram
additional information
?
-
-
cyclization of (2Z,6E)-farnesyl diphosphate by Cop6 proceeds through opposite enantiomers of a beta-bisabolyl cation intermediate and results in alpha-acoradiene + amorpha-4,11-diene
-
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0076 - 0.04
(2E,6E)-farnesyl diphosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.67
(2E,6E)-farnesyl diphosphate
Coprinopsis cinerea
-
at 25C, pH 8.0; at 25C, pH not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.246 - 88
(2E,6E)-farnesyl diphosphate
81
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Talon resin column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli JM109 cells and in Saccharomyces cerevisiae strain YPH 500
-
expression in Escherichia coli, Saccharomyces cerevisiae and Xanthophyllomyces dendrorhous
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C236A
-
the mutant produces 96% (-)-alpha-cuprenene, 3% gamma-cuprenene and 1% sesquiterpene products from (2E,6E)-farnesyl diphosphate (41.2% of wild type activity)
E237L
-
the mutant produces 96% (-)-alpha-cuprenene, 3% gamma-cuprenene and 1% unidentified sesquiterpene products from (2E,6E)-farnesyl diphosphate (59.4% of wild type activity)
N224D
-
the mutant makes a small amount of (6S)-beta-bisabolene (18%) from (2Z,6E)-farnesyl diphosphate, the mutation does not affect the product profile of Cop6 with its normal substrate (2E,6E)-farnesyl diphosphate
N240L
-
the mutant produces 98% (-)-alpha-cuprenene and 2% gamma-cuprenene from (2E,6E)-farnesyl diphosphate (31% of wild type activity)
additional information
-
in the Cop6 loop graft mutant Cop6L4, loop replacement has no influence on the selectivity of the enzyme for alpha-cuprenene, the mutation reduces the catalytic efficiency 3-17fold
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
expression in Xanthophyllomyces dendrorhous leads to production of alpha-cuprenene up to 80 mg/l. At this expression levels the pool of terpene precursors is not a limiting factor since the expression of the Cop6 gene in the genomic rDNA of the yeast allows production of both alpha-cuprenene and astaxanthin without affecting the growth or the accumulation levels of both compounds