BRENDA - Enzyme Database show
show all sequences of 1.14.12.23

Protein engineering of the archetypal nitroarene dioxygenase of Ralstonia sp. strain U2 for activity on aminonitrotoluenes and dinitrotoluenes through alpha-subunit residues leucine 225, phenylalanine 350, and glycine 407

Keenan, B.G.; Leungsakul, T.; Smets, B.F.; Mori, M.A.; Henderson, D.E.; Wood, T.K.; J. Bacteriol. 187, 3302-3310 (2005)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
F350T
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
Ralstonia sp.
F350T/G407S
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
Ralstonia sp.
G50S/L225R/A269T
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
Ralstonia sp.
L225R
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
Ralstonia sp.
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Ralstonia sp.
-
-
-
Ralstonia sp. U2
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,3-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
4-methyl-3-nitrocatechol + nitrite + NAD+
-
-
-
?
2,3-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
4-methyl-3-nitrocatechol + nitrite + NAD+
-
-
-
?
2,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
3-methyl-4-nitrocatechol + nitrite + NAD+
-
-
-
?
2,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
3-methyl-4-nitrocatechol + nitrite + NAD+
-
-
-
?
2-amino-4,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
3-amino-4-methyl-5-nitrocatechol + nitrite + NAD+
plus 2-amino-4,6-dinitrobenzyl alcohol
-
-
?
2-amino-4,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
3-amino-4-methyl-5-nitrocatechol + nitrite + NAD+
plus 2-amino-4,6-dinitrobenzyl alcohol
-
-
?
additional information
wild-type enzyme catalyzes dioxygenation of naphthalen, reaction of EC 1.14.12.12
734066
Ralstonia sp.
?
-
-
-
-
additional information
wild-type enzyme catalyzes dioxygenation of naphthalen, reaction of EC 1.14.12.12
734066
Ralstonia sp. U2
?
-
-
-
-
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
F350T
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
Ralstonia sp.
F350T/G407S
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
Ralstonia sp.
G50S/L225R/A269T
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
Ralstonia sp.
L225R
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
Ralstonia sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,3-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
4-methyl-3-nitrocatechol + nitrite + NAD+
-
-
-
?
2,3-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
4-methyl-3-nitrocatechol + nitrite + NAD+
-
-
-
?
2,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
3-methyl-4-nitrocatechol + nitrite + NAD+
-
-
-
?
2,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
3-methyl-4-nitrocatechol + nitrite + NAD+
-
-
-
?
2-amino-4,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp.
3-amino-4-methyl-5-nitrocatechol + nitrite + NAD+
plus 2-amino-4,6-dinitrobenzyl alcohol
-
-
?
2-amino-4,6-dinitrotoluene + NADH + O2
no substrate for wild-type, but substrate of mutant F350T
734066
Ralstonia sp. U2
3-amino-4-methyl-5-nitrocatechol + nitrite + NAD+
plus 2-amino-4,6-dinitrobenzyl alcohol
-
-
?
additional information
wild-type enzyme catalyzes dioxygenation of naphthalen, reaction of EC 1.14.12.12
734066
Ralstonia sp.
?
-
-
-
-
additional information
wild-type enzyme catalyzes dioxygenation of naphthalen, reaction of EC 1.14.12.12
734066
Ralstonia sp. U2
?
-
-
-
-
Other publictions for EC 1.14.12.23
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733176
Mahan
Selection for growth on 3-nitr ...
Acidovorax sp., Acidovorax sp. JS42
Appl. Environ. Microbiol.
81
309-319
2015
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-
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-
4
-
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18
-
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3
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12
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17
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4
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18
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12
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17
-
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-
1
1
-
18
18
733264
Singh
Expression, purification and s ...
Diaphorobacter sp. DS2
Biochem. Biophys. Res. Commun.
445
36-42
2014
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-
1
-
1
-
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-
1
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5
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1
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1
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5
-
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-
733159
Liu
Patchwork assembly of nag-like ...
Pseudomonas stutzeri ZWLR2-1
Appl. Environ. Microbiol.
77
4547-4552
2011
-
-
-
-
-
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5
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1
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1
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1
1
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733154
Ju
Control of substrate specifici ...
Comamonas sp.
Appl. Environ. Microbiol.
72
1817-1824
2006
-
-
1
-
6
-
-
10
-
-
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1
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4
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1
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6
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10
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4
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733153
Parales
Purification, characterization ...
Acidovorax sp., Acidovorax sp. JS42
Appl. Environ. Microbiol.
71
3806-3814
2005
-
-
-
1
-
-
-
2
-
-
7
-
-
3
-
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-
-
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4
1
-
-
-
2
-
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-
2
-
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2
1
-
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-
2
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7
-
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4
1
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-
2
-
-
-
-
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-
-
-
-
-
734066
Keenan
Protein engineering of the arc ...
Ralstonia sp., Ralstonia sp. U2
J. Bacteriol.
187
3302-3310
2005
-
-
-
-
4
-
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6
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8
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4
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8
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734469
Friemann
Structural insight into the di ...
Comamonas sp., Comamonas sp. JS7650
J. Mol. Biol.
348
1139-1151
2005
-
-
-
1
-
-
-
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-
1
-
-
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2
-
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1
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1
1
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1
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-
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-
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-
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-
733150
Lessner
Molecular characterization and ...
Comamonas sp.
Appl. Environ. Microbiol.
68
634-641
2002
-
-
-
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4
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7
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7
-
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734057
Parales
Enzyme specificity of 2-nitrot ...
Pseudomonas sp., Pseudomonas sp. JS42
J. Bacteriol.
180
1194-1199
1998
-
-
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1
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5
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8
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1
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8
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