Information on EC 4.2.3.37 - epi-isozizaene synthase

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The expected taxonomic range for this enzyme is: Streptomyces

EC NUMBER
COMMENTARY hide
4.2.3.37
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RECOMMENDED NAME
GeneOntology No.
epi-isozizaene synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2E,6E)-farnesyl diphosphate = (+)-epi-isozizaene + diphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cyclization
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
albaflavenone biosynthesis
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Sesquiterpenoid and triterpenoid biosynthesis
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Biosynthesis of secondary metabolites
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Biosynthesis of antibiotics
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SYSTEMATIC NAME
IUBMB Comments
(2E,6E)-farnesyl-diphosphate diphosphate-lyase [(+)-epi-isozizaene-forming]
Requires Mg2+ for activity. The displacement of the diphosphate group of farnesyl diphosphate occurs with retention of configuration [1]. In the soil-dwelling bacterium Streptomyces coelicolor A3(2), the product of this reaction is used by EC 1.14.13.106, epi-isozizaene 5-monooxygenase, to produce the sesquiterpene antibiotic albaflavenone [2].
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
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
(+)-epi-isozizaene + diphosphate
show the reaction diagram
(3R)-(Z)-nerolidyl diphosphate
(+)-epi-isozizaene + diphosphate
show the reaction diagram
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
(+)-epi-isozizaene + diphosphate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
32% of the activity with Mg2+
Zn2+
5% of the activity with Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
complete inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000125 - 0.00145
(2E,6E)-farnesyl diphosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00024 - 0.049
(2E,6E)-farnesyl diphosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.23 - 63
(2E,6E)-farnesyl diphosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
recombinant mutants F96S and F96N, pH 7.5, 30C
4.2
recombinant mutant W230H, pH 7.5, 30C
5
recombinant mutants F95N and Y69A, pH 7.5, 30C
5.5
recombinant mutant F95C, pH 7.5, 30C
5.8
recombinant mutant F96H, pH 7.5, 30C
5.9
recombinant mutant F96T, pH 7.5, 30C
5.92
recombinant mutant F95Q, pH 7.5, 30C
6
recombinant mutant W230Y, pH 7.5, 30C
7
recombinant mutant F96M, pH 7.5, 30C
7.6
recombinant mutant Y69F, pH 7.5, 30C
8
recombinant mutants F96Q and F95Y, pH 7.5, 30C
18
recombinant wild-type enzyme, pH 7.5, 30C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified wild-type enzyme and mutant F95N in complex with Mg2+, benzyl triethylammonium cation (BTAC), and diphosphate, sitting drop vapor diffusion method, mixing of 5 mm nl of 5 mg/ml protein in 300 mM NaCl, 20 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 2 mM tris(2-carboxyethyl)phosphine (TCEP), 10% glycerol, 2 mM sodium pyrophosphate, and 2 mM BTAC with 600 nl of precipitant solution containing 0.17 M sodium acetate trihydrate, 85 mM sodium cacodylate trihydrate, pH 6.5, 25.5% PEG 8000, and 15% glycerol, addition of 100 nL of the 100fold dilution of wild-type enzyme crystallization seed stock, equilibration against a 0.1 ml reservoir of the precipitant solution at room temperature, 6 days, X-ray diffraction structure determination and analysis at 1.8 A and 2.51 A resolution, respectively
sitting drop vapor diffusion method
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
immobilized metal ion affinity chromatography (Co2+)
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
His-tagged version expressed in Escherichia coli BL21(DE3)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D100N
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below 5% of wild-type activity
D99E
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below 5% of wild-type activity
E248D
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below 5% of wild-type activity
F198A
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major products: Z-gamma-bisabolene (24%), sesqusabine (20%)
F95C
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F95N
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F95Q
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F95Y
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F96A
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major product: beta-farnesene (70%)
F96H
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F96M
site-directed mutagenesis, the substitution converts the epi-isozizaene synthase into a high-fidelity sesquisabinene synthase, the mutant generates 91% sesquisabinene A
F96N
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
F96Q
site-directed mutagenesis, the substitution converts the epi-isozizaene synthase into a high-fidelity sesquisabinene synthase, the mutant generates 97% sesquisabinene A
F96S
site-directed mutagenesis, the substitution converts the epi-isozizaene synthase into a high-fidelity sesquisabinene synthase, the mutant generates 78% sesquisabinene A
F96T
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
N240D
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below 5% of wild-type activity
S244A
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below 5% of wild-type activity
W203F
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major product: Z-gamma-bisabolene (47%)
W203H
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
W203Y
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
Y69A
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
Y69F
site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
D100N
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below 5% of wild-type activity
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D99E
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below 5% of wild-type activity
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D99N
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completely inactive
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F96S
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site-directed mutagenesis, the substitution converts the epi-isozizaene synthase into a high-fidelity sesquisabinene synthase, the mutant generates 78% sesquisabinene A
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N240D
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below 5% of wild-type activity
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S244A
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below 5% of wild-type activity
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W203H
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site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
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Y69A
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site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
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Y69F
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site-directed mutagenesis, the mutant shows reduced activity with farnesyl diphosphate compared to the wild-type
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additional information