Information on EC 1.14.12.B2 - 2-nitrotoluene 2,3-dioxygenase [NADH]

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.12.B2
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
2-nitrotoluene 2,3-dioxygenase [NADH]
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SYSTEMATIC NAME
IUBMB Comments
2-nitrotoluene,NADH:oxygen oxidoreductase (1,2-hydroxylating, denitrifying)
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
M9PTH9 i.e. ferredoxin reductase subunit MntAa, M9PW10 i.e. oxygenase alpha subunit MntAc, M9PSS0 i.e. ferredoxin subunit MntAb, M9PV03 i.e. oxygenase beta subunit MntAd
M9PTH9 and M9PW10 and M9PSS0 and M9PV03
UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
2,3-dinitrotoluene + NADH + O2
4-methyl-3-nitrocatechol + nitrite + NAD+
show the reaction diagram
2,6-dinitrotoluene + NADH + O2
3-methyl-4-nitrocatechol + nitrite + NAD+
show the reaction diagram
2-amino-4,6-dinitrotoluenedinitrotoluene + NADH + O2
3-amino-4-methyl-5-nitrocatechol + nitrite + NAD+
show the reaction diagram
2-chloronitrobenzene + NADH + O2
3-chlorocatechol + nitrite + NAD+
show the reaction diagram
M9PTH9 and M9PW10 and M9PSS0 and M9PV03
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80% of the activity with 3-nitrotoluene
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?
2-nitrotoluene + NADH + O2
3-methylcatechol + nitrite + NAD+
show the reaction diagram
3-nitrotoluene + NADH + O2
3-methylcatechol + 4-methylcatechol + nitrite + NAD+
show the reaction diagram
M9PTH9 and M9PW10 and M9PSS0 and M9PV03
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products are 54% 3-methylcatechol, 39% 4-methylcatechol, 7% 3-nitrobenzyl alcohol
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?
3-nitrotoluene + NADH + O2
3-methylcatechol + nitrite + NAD+
show the reaction diagram
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products are 72% 3-methylcatechol, 25% 4-methylcatechol and 4% 3-nitrobenzyl alcohol
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?
3-nitrotoluene + NADH + O2
4-methylcatechol + nitrite + NAD+
show the reaction diagram
4-nitrotoluene + NADH + O2
4-methylcatechol + nitrite + NAD+
show the reaction diagram
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?
naphthalene + NADH + O2
? + nitrite + NAD+
show the reaction diagram
naphthalene + O2
cis-1,2-dihydroxy-1,2-dihydronaphthalene
show the reaction diagram
nitrobenzene + NADH + O2
catechol + nitrite + NAD+
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
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0.5 mol of NADH is required to reduce each alphabeta heterodimer of oxygenase
[2Fe-2S]-center
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presence of a Rieske [2Fe-2S] cluster in the oxygenase component
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0012 - 0.02
2-nitrotoluene
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0.0039 - 0.04
3-nitrotoluene
0.121
Naphthalene
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pH 6.8, 30C
0.0047 - 0.047
nitrobenzene
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12 - 2.3
2-nitrotoluene
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0.13 - 0.27
3-nitrotoluene
2.4
Naphthalene
Acidovorax sp.
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pH 6.8, 30C
0.19 - 0.27
nitrobenzene
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.5 - 160
2-nitrotoluene
206831
3.7 - 68
3-nitrotoluene
136150
4 - 58
nitrobenzene
21873
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25500
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3 * 50400, alpha-subunit, 3 * 25500, beta-subunit, SDS-PAGE of oxygenase component
50400
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3 * 50400, alpha-subunit, 3 * 25500, beta-subunit, SDS-PAGE of oxygenase component
210000
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gel filtration, oxygenase component
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
oxygenase component, to 1.2 A resolution, hexagonal space group P63 with cell dimensions a = b = 121.6 A and c = 84.4 A
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
M9PTH9 and M9PW10 and M9PSS0 and M9PV03
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A405G
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synthesizes 17% 3-methylcatechol, 83% 4-methylcatechol from substrate 3-nitrotoluene
I204A
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synthesizes 2% 3-methylcatechol, 97% 4-methylcatechol and 1% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
I204T
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synthesizes 5% 3-methylcatechol, 93% 4-methylcatechol and 2% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
I204V
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synthesizes 13% 3-methylcatechol, 83% 4-methylcatechol and 5% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
A405G
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synthesizes 17% 3-methylcatechol, 83% 4-methylcatechol from substrate 3-nitrotoluene
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I204A
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synthesizes 2% 3-methylcatechol, 97% 4-methylcatechol and 1% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
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I204T
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synthesizes 5% 3-methylcatechol, 93% 4-methylcatechol and 2% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
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I204V
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synthesizes 13% 3-methylcatechol, 83% 4-methylcatechol and 5% 3-nitrobenzyl alcohol from substrate 3-nitrotoluene
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F350T
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contrary to wild-type, mutant produces 3-methyl-4-nitrocatechol from 2,6-dinitrotoluene and 3-amino-4-methyl-5-nitrocatechol and 2-amino-4,6-dinitrobenzyl alcohol from 2-amino-4,6-dinitrotoluene, and mutant releases nitrite from 2,3-dinitrotoluene sixfold faster than wild-type
F350T/G407S
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double mutant increases the rate of oxidation of 2,3-dinitrotoluene, 2,6-dinitrotoluene , and 2-amino-4,6-dinitrotoluene threefold relative to variant F350T
G50S/L225R/A269T
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mutant displays increased rate of 4-amino-2-nitrotoluene oxidation and additionally produces oxidation product 4-amino-2-nitrocresol (enhanced 11fold relative to wild-type) as well as 4-amino-2-nitrobenzyl alcohol
L225R
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12fold faster generation of 4-amino-2-nitrocresol and production of 4-amino-2-nitrobenzyl alcohol from 4-amino-2-nitrotoluene as well as 24fold faster generation of nitrite and 15fold faster generation of 2,3-dinitrobenzyl alcohol from 2,3-dinitrotoluene
F350T
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contrary to wild-type, mutant produces 3-methyl-4-nitrocatechol from 2,6-dinitrotoluene and 3-amino-4-methyl-5-nitrocatechol and 2-amino-4,6-dinitrobenzyl alcohol from 2-amino-4,6-dinitrotoluene, and mutant releases nitrite from 2,3-dinitrotoluene sixfold faster than wild-type
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F350T/G407S
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double mutant increases the rate of oxidation of 2,3-dinitrotoluene, 2,6-dinitrotoluene , and 2-amino-4,6-dinitrotoluene threefold relative to variant F350T
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G50S/L225R/A269T
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mutant displays increased rate of 4-amino-2-nitrotoluene oxidation and additionally produces oxidation product 4-amino-2-nitrocresol (enhanced 11fold relative to wild-type) as well as 4-amino-2-nitrobenzyl alcohol
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L225R
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12fold faster generation of 4-amino-2-nitrocresol and production of 4-amino-2-nitrobenzyl alcohol from 4-amino-2-nitrotoluene as well as 24fold faster generation of nitrite and 15fold faster generation of 2,3-dinitrobenzyl alcohol from 2,3-dinitrotoluene
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additional information