Information on EC 4.2.3.22 - germacradienol synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
4.2.3.22
-
RECOMMENDED NAME
GeneOntology No.
germacradienol synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2E,6E)-farnesyl diphosphate + H2O = (1E,4S,5E,7R)-germacra-1(10),5-dien-11-ol + diphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
additional information
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
-
geosmin biosynthesis
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-
Metabolic pathways
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Sesquiterpenoid and triterpenoid biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
(2E,6E)-farnesyl-diphosphate diphosphate-lyase [(1E,4S,5E,7R)-germacra-1(10),5-dien-11-ol-forming]
Requires Mg2+ for activity. H-1si of farnesyl diphosphate is lost in the formation of (1E,4S,5E,7R)-germacra-1(10),5-dien-11-ol. Formation of (-)-germacrene D involves a stereospecific 1,3-hydride shift of H-1si of farnesyl diphosphate. Both products are formed from a common intermediate [2]. Other enzymes produce germacrene D as the sole product using a different mechanism. The enzyme mediates a key step in the biosynthesis of geosmin (see EC 4.1.99.16 geosmin synthase), a widely occurring metabolite of many streptomycetes, bacteria and fungi [2]. Also catalyses the reaction of EC 4.2.3.75, (-)-germacrene D synthase.
CAS REGISTRY NUMBER
COMMENTARY hide
211049-88-6
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
gene SC9B1.20
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-
Manually annotated by BRENDA team
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 + H2O
(1E,4S,5E,7R)-germa-1(10),5-dien-11-ol + diphosphate
show the reaction diagram
-
catalysed by the N-terminal domain of the bifunctional enzyme
-
?
(2E,6E)-farnesyl diphosphate + H2O
(1E,4S,5E,7R)-germacra-1(10),5-dien-11-ol + diphosphate
show the reaction diagram
2-trans,6-trans-farnesyl diphosphate + H2O
(4S,7R)-germacra-1(10)E,5E-dien-11-ol + diphosphate
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
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 + H2O
(1E,4S,5E,7R)-germacra-1(10),5-dien-11-ol + diphosphate
show the reaction diagram
2-trans,6-trans-farnesyl diphosphate + H2O
(4S,7R)-germacra-1(10)E,5E-dien-11-ol + diphosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe3+
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can substitute for Mg2+
Ni2+
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can substitute for Mg2+
additional information
-
enzyme activity is dependent on divalent cations
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
suppresses the formation of geosmin by factor 3 or more
Fe2+
suppresses the formation of geosmin by factor 3 or more
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000062
(2E,6E)-farnesyl diphosphate
0.000115
2-trans,6-trans-farnesyl diphosphate
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pH 8.2, 30C, recombinant N-terminal domain
0.000075
farnesyl diphosphate
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0062
(2E,6E)-farnesyl diphosphate
Streptomyces coelicolor
Q9X839
pH 8.2, 30C
3.2 - 6.2
2-trans,6-trans-farnesyl diphosphate
0.0031
farnesyl diphosphate
Streptomyces avermitilis
Q82L49
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
100
(2E,6E)-farnesyl diphosphate
Streptomyces coelicolor
Q9X839
pH 8.2, 30C
81
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
activity assay
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
strain ATCC 27952 produces clinically important anthracycline chemotherapeutic agents of the polyketide class of antibiotics, daunorubicin and doxorubicin
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
83000
determined by SDS-PAGE
98000
recombinant Spterp13, consistent with the predicted mass of Spterp13 plus his-tag determined by SDS-PAGE analysis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
the recombinant N-terminal half of the protein catalyzes the Mg2+-dependent cyclization of farnesyl diphosphate to germacradienol and germacrene D, while the highly homologous C-terminal domain catalyzes the Mg2+-dependent conversion of germacradienol to geosmin
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purification by Co2+-affinity chromatography
recombinant enzyme from Escherichia coli strain BL21(DE3), soluble enzyme and enzyme solubilized and refolded from inclusion bodies, by ion exchange chromatography and gel filtration
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using Q-Sepharose resin and a Q-Sepharose column
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of an ordered library of Supercos-1 clones, DNA sequence determination and analysis, subcloning in Escherichia coli
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expression in Escherichia coli strain BL21(DE3)
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for construction of a geoA-deletion mutant of Streptomyces avermitilis, and for expression in Escherichia coli BL21DE3 cells
for expression in Escherichia coli
full length recombinant protein is heterologously expressed as a his-tagged fusion protein in Escherichia coli
into the pCR2.1-TOPO vector, for functional expression full-length cDNA is sub-cloned into a pET vector
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
resuspended inclusion body protein is refolded
solubilization of recombinant enzyme from inclusion bodies by 0.02% Triton X-100 and 100 mM NaOH, and refolding
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