Information on EC 1.13.12.19 - 2-oxoglutarate dioxygenase (ethylene-forming)

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The expected taxonomic range for this enzyme is: Pseudomonas syringae

EC NUMBER
COMMENTARY
1.13.12.19
-
RECOMMENDED NAME
GeneOntology No.
2-oxoglutarate dioxygenase (ethylene-forming)
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2-oxoglutarate + O2 = ethylene + 3 CO2 + H2O
show the reaction diagram
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
ethylene biosynthesis II (microbes)
-
ethylene biosynthesis IV
-
ethylene biosynthesis V (engineered)
-
ethylene forming enzyme
-
SYSTEMATIC NAME
IUBMB Comments
2-oxoglutarate:oxygen oxidoreductase (decarboxylating, ethylene-forming)
This is one of two simultaneous reactions catalysed by the enzyme, which is responsible for ethylene production in bacteria of the Pseudomonas syringae group. In the other reaction [EC 1.14.11.34, 2-oxoglutarate/L-arginine monooxygenase/decarboxylase (succinate-forming)] the enzyme catalyses the mono-oxygenation of both 2-oxoglutarate and L-arginine, forming succinate, carbon dioxide and L-hydroxyarginine, which is subsequently cleaved into guanidine and (S)-1-pyrroline-5-carboxylate. The enzymes catalyse two cycles of the ethylene-forming reaction for each cycle of the succinate-forming reaction, so that the stoichiometry of the products ethylene and succinate is 2:1.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
ethylene forming enzyme
-
-
ethylene-forming enzyme
P32021
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
gene is encoded by an indigenous plasmid, designated pPSP1; pv. phaseolicola PK2
UniProt
Manually annotated by BRENDA team
pv. phaseolicola PK2
UniProt
Manually annotated by BRENDA team
pv. phaseolicola PK2, enzyme catalyzes the formation of ethylene and succinate from 2-oxoglutarate, reactions of EC 1.13.12.19 and EC 1.14.11.34
UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-oxoglutarate + O2
ethylene + 3 CO2 + H2O
show the reaction diagram
-
-
-
-
?
2-oxoglutarate + O2
ethylene + 3 CO2 + H2O
show the reaction diagram
P32021
enzyme is highly specific for substrate 2-oxoglutarate
-
-
?
2-oxoglutarate + O2
ethylene + ?
show the reaction diagram
P32021
presence of oxygen is essential for the ethylene forming reaction by EFE
-
-
?
additional information
?
-
P32021
enzyme catalyzes the formation of ethylene and succinate from 2-oxoglutarate, at a molar ratio of 2:l, reactions of EC 1.13.12.19 and EC 1.14.11.34. In the main reaction, 2-oxoglutarate is dioxygenated to produce one molecule of ethylene and three molecules of carbon dioxide. In the sub-reaction, both 2-oxoglutarate and L-arginine are mono-oxygenated to yield succinate plus carbon dioxide and L-hydroxyarginine, respectively, the latter being further transformed to guanidine and L-delta-pyrroline-5-carboxylate. Dual-circuit mechanism for the entire reaction is proposed, in which the binding of L-arginine and 2-oxoglutarate in a Schiff-base structure generates a common intermediate for two reactions
-
-
-
additional information
?
-
P32021
presence of 2-oxoglutarate, L-arginine, Fe2+ and oxygen is essential for the enzymic reaction
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
at the active site, enzyme acts as a bidentate ligand and it forms a complex with Fe2+. The Fe2+ is further coordinated to a tridentate Schiff base of 2-oxoglutarate and L-arginine, whose terminal carboxylate and guanidino groups are trapped by binding sites I and II on the enzyme, respectively
Fe2+
P32021
required, KM value 0.059 mM
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4,5-Dihydroxy-1,3-benzene disulfonic acid
P32021
1 mM, 0.8% residual activity
5,5'-dithio-bis(2-nitrobenzoate)
P32021
1 mM, 0.7% residual activity
CoCl2
P32021
1 mM, 20% residual activity
CuSO4
P32021
1 mM, 50% residual activity
EDTA
P32021
1 mM, 1% residual activity
H2O2
P32021
1 mM, 0.7% residual activity
MnCl2
P32021
1 mM, 6% residual activity
n-propyl gallate
P32021
1 mM, 1% residual activity
Sodium azide
P32021
1 mM, 90% residual activity
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
D-Arginine
P32021
3% of the activity with L-arginine
L-arginine
P32021
highly specific for cofactor L-arginine, KM value 0.018 mM
L-canavanine sulfate
P32021
7% of the activity with L-arginine
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.006
-
2-oxoglutarate
-
mutant H305Q, pH 8.0, 25C
0.007
-
2-oxoglutarate
-
mutant H168Q, pH 8.0, 25C; mutant H284Q, pH 8.0, 25C
0.008
-
2-oxoglutarate
-
mutant H335Q, pH 8.0, 25C
0.01
-
2-oxoglutarate
-
mutant H309Q, pH 8.0, 25C
0.011
-
2-oxoglutarate
-
mutant H169Q, pH 8.0, 25C
0.013
-
2-oxoglutarate
-
wild-type, pH 8.0, 25C
0.015
-
2-oxoglutarate
-
mutant H116Q, pH 8.0, 25C
0.019
-
2-oxoglutarate
P32021
pH 8.0, 25C
0.033
-
2-oxoglutarate
-
mutant H268Q, pH 8.0, 25C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.009
-
2-oxoglutarate
-
mutant H268Q, pH 8.0, 25C
0.01
-
2-oxoglutarate
-
mutant H284Q, pH 8.0, 25C
0.012
-
2-oxoglutarate
-
mutant H116Q, pH 8.0, 25C
0.015
-
2-oxoglutarate
-
mutant H168Q, pH 8.0, 25C
0.017
-
2-oxoglutarate
-
mutant H309Q, pH 8.0, 25C
0.047
-
2-oxoglutarate
-
mutant H169Q, pH 8.0, 25C
0.2
-
2-oxoglutarate
-
mutant H305Q, pH 8.0, 25C
0.3
-
2-oxoglutarate
-
mutant H335Q, pH 8.0, 25C
0.5
-
2-oxoglutarate
-
wild-type, pH 8.0, 25C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.27
-
2-oxoglutarate
-
mutant H268Q, pH 8.0, 25C
34
0.8
-
2-oxoglutarate
-
mutant H116Q, pH 8.0, 25C
34
1.43
-
2-oxoglutarate
-
mutant H284Q, pH 8.0, 25C
34
1.67
-
2-oxoglutarate
-
mutant H309Q, pH 8.0, 25C
34
2.14
-
2-oxoglutarate
-
mutant H168Q, pH 8.0, 25C
34
4.24
-
2-oxoglutarate
-
mutant H169Q, pH 8.0, 25C
34
33.3
-
2-oxoglutarate
-
mutant H305Q, pH 8.0, 25C
34
37.5
-
2-oxoglutarate
-
mutant H335Q, pH 8.0, 25C
34
38.5
-
2-oxoglutarate
-
wild-type, pH 8.0, 25C
34
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
660
-
P32021
pH 8.0, 25C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
7.5
P32021
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
25
P32021
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.9
-
P32021
isoelectric focusing
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
36000
-
P32021
gel filtration
36000
-
P32021
gel fitlration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
monomer
P32021
1 * 39444, calculated, 1 * 42000, SDS-PAGE
monomer
P32021
1 * 42000, SDS-PAGE
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
P32021
stable below
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
from cell-free extract
P32021
recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Saccharomyces cerevisae. Different cultivation factors on ethylene formation in Saccharomyces cerevisiae expressing the EFE in continuous cultures are investigated. Main finding is that oxygen availability is crucial for ethylene production. By employing three different nitrogen sources it is shown that the nitrogen source available can both improve and impair the ethylene productivity. N-Source/yield ethylene (microgram/g glucose): (NH4)2SO4/164, glutamate/233, glutamate+arginine/96.8
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expression in Escherichia coli
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expression in Nicotiana tabacum
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
H116Q
-
kcat value decreases to 2.4% of wild-type. Mutant is more thermolabile than wild-type
H168Q
-
kcat value decreases to 3% of wild-type. Mutant is more thermolabile than wild-type
H169Q
-
kcat value decreases to 9.3% of wild-type. Mutant is more thermolabile than wild-type
H189Q
-
complete loss of activity
H233Q
-
complete loss of activity
H268Q
-
kcat value decreases to 1.8% of wild-type
H284Q
-
kcat value decreases to 2% of wild-type. Mutant is more thermolabile than wild-type
H305Q
-
kcat value decreases to 40% of wild-type
H309Q
-
kcat value decreases to 3.3% of wild-type. Mutant is more thermolabile than wild-type
H335Q
-
kcat value decreases to 60% of wild-type
additional information
-
introduction of a gene encoding a chimeric protein consisting of EFE and beta-glucuronidase GUS into the tobacco genome using a binary vector which directs expression of the EFE-GUS fusion protein under the control of constitutive promoter of cauliflower mosaic virus 35S RNA
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
agriculture
-
introduction of a gene encoding a chimeric protein consisting of EFE and beta-glucuronidase GUS into the tobacco genome using a binary vector which directs expression of the EFE-beta-glucuronidase fusion protein under the control of constitutive promoter of cauliflower mosaic virus 35S RNA. Transgenic plants produce ethylene at consistently higher rates than the untransformed plant, and their beta-glucuronidase activities are expressed in different tissues. A significant dwarf morphology observed in the transgenic tobacco displaying the highest ethylene production resembles the phenotype of a wild-type plant exposed to excess ethylene
biotechnology
-
different cultivation factors on ethylene formation in Saccharomyces cerevisiae expressing the EFE in continuous cultures is investigated. Main finding is that oxygen availability is crucial for ethylene production. By employing three different nitrogen sources it is shown that the nitrogen source available can both improve and impair the ethylene productivity