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Information on EC 5.3.2.6 - 2-hydroxymuconate tautomerase and Organism(s) Pseudomonas putida and UniProt Accession Q01468
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5.3.2.6 2-hydroxymuconate tautomeraseIUBMB Comments
Involved in the meta-cleavage pathway for the degradation of phenols, modified phenols and catechols. The enol form (2Z,4E)-2-hydroxyhexa-2,4-dienedioate is produced as part of this pathway and is converted to the keto form (3E)-2-oxohex-3-enedioate by the enzyme . Another keto form, (4E)-2-oxohex-4-enedioate (4-oxalocrotonate), was originally thought to be produced by the enzyme [1,2] but later shown to be produced non-enzymically .
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Word Map
- 5.3.2.6
-
tautomerization
-
michael-type
-
trans-3-chloroacrylic
- 5-carboxymethyl-2-hydroxymuconate
- 2-oxo-3-pentynoate
-
stivers
-
dienolic
-
heterohexamer
- 2-hydroxy-2,4-pentadienoate
- 3-bromopyruvate
-
beta-alpha-beta
-
whitman
-
proline-based
-
3-chloroacrylic
- cis-caad
-
meta-fission
-
cis-3-chloroacrylic
The taxonomic range for the selected organisms is: Pseudomonas putida
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
4-oxalocrotonate tautomerase, afdmpi, hpdmpi, sar1376, 2-hydroxymuconate tautomerase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
4-OT
-
-
4-oxalocrotonate tautomerase
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate = (3E)-2-oxohex-3-enedioate
catalytic reaction mechanism, detailed overview
-
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate = (3E)-2-oxohex-3-enedioate
catalytic reaction mechanism, detailed overview. Quantum mechanics/molecular mechanics simulation of proton transfer reaction and structure-function analysis, the ketonization mechanism in 4-OT depends on the enzymatic model used, detailed overview
-
PATHWAY SOURCE
PATHWAYS
KEGG
-
-, -, -, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate keto-enol isomerase
Involved in the meta-cleavage pathway for the degradation of phenols, modified phenols and catechols. The enol form (2Z,4E)-2-hydroxyhexa-2,4-dienedioate is produced as part of this pathway and is converted to the keto form (3E)-2-oxohex-3-enedioate by the enzyme [6]. Another keto form, (4E)-2-oxohex-4-enedioate (4-oxalocrotonate), was originally thought to be produced by the enzyme [1,2] but later shown to be produced non-enzymically [5].
CAS REGISTRY NUMBER
COMMENTARY
85876-28-4
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate
(3E)-2-oxohex-3-enedioate
-
-
-
r
acetaldehyde + (1Z)-4-methyl-1-nitropent-1-ene
6-methyl-4-(nitromethyl)heptan-2-one
-
-
-
r
acetaldehyde + 1-chloro-4-[(E)-2-nitroethenyl]benzene
4-(4-chlorophenyl)-5-nitropentan-2-one
-
-
-
r
acetaldehyde + 1-fluoro-4-[(E)-2-nitroethenyl]benzene
4-(4-fluorophenyl)-5-nitropentan-2-one
-
-
-
r
acetaldehyde + 2-(cyclopentyloxy)-1-methoxy-4-[(E)-2-nitroethenyl]benzene
4-[3-(cyclopentyloxy)-4-methoxyphenyl]-5-nitropentan-2-one
-
-
-
r
acetaldehyde + 2-methoxy-5-[(E)-2-nitroethenyl]phenol
4-(3-hydroxy-4-methoxyphenyl)-5-nitropentan-2-one
-
-
-
r
acetaldehyde + 4-[(E)-2-nitroethenyl]phenol
4-(4-hydroxyphenyl)-5-nitropentan-2-one
-
-
-
r
acetaldehyde + [(E)-2-nitroethenyl]benzene
5-nitro-4-phenylpentan-2-one
-
-
-
r
butanal + [(E)-2-nitroethenyl]benzene
3-ethyl-5-nitro-4-phenylpentan-2-one
-
-
-
r
(E)-1-nitro-2-(2-thienyl)ethene + isobutanal
2,2-dimethyl-4-nitro-3-(thiophen-2-yl)butanal
-
-
-
-
?
(E)-2-(furan-2-yl)nitroethene + isobutanal
3-(furan-2-yl)-2,2-dimethyl-4-nitrobutanal
-
-
-
-
?
(E)-2-(thiophen-2-yl)nitroethene + isobutanal
2,2-dimethyl-4-nitro-3-(thiophen-2-yl)butanal
-
-
-
-
?
2-chloro-beta-nitrostyrene + acetaldehyde
3-(2-chlorophenyl)-4-nitrobutanal
-
-
51% yield
-
?
2-hydroxy-2,4-hexadienedioate
2-oxo-3,4-hexenedioate
-
4-OT with 2-hydroxy-2,4-hexadienedioate in D2O results in a racemic mixture of 2-oxo-[3-2H]-4-hexenedioate, suggesting that 4-OT may not catalyze a 1,3-keto-enol tautomerization reaction using this dienol
-
-
?
2-hydroxymuconate
2-oxo-3-(E)-hexenedioate
-
stereospecific ketonization
-
-
?
2-oxo-4(E)-hexenedioate
2-oxo-3(E)-hexenedioate
-
1,3-allylic isomerization
-
-
?
3,4-(methylenedioxy)-beta-nitrostyrene + isobutanal
(3E)-3-(2H-1,3-benzodioxol-5-yl)-2,2-dimethyl-4-nitrobut-3-enal
-
-
-
-
?
4-chloro-beta-nitrostyrene + acetaldehyde
3-(4-chlorophenyl)-4-nitrobutanal
-
-
38% yield
-
?
4-fluoro-beta-nitrostyrene + acetaldehyde
3-(4-fluorophenyl)-4-nitrobutanal
-
-
31% yield
-
?
4-vinyl-2,3-dihydropyrrole-2-carboxylic acid
4-ethylidene-3,4-dihydropyrrole-2-carboxylic acid
-
-
-
-
r
beta-nitrostyrene + acetaldehyde
4-nitro-3-phenyl-butanal
-
-
60% yield
-
?
beta-nitrostyrene + isobutanal
2,2-dimethyl-4-nitro-3-phenylbutanal
-
-
-
-
?
isobutanal + beta-nitrostyrene
2,2-dimethyl-4-nitro-3-phenylbutanal + 4-nitro-3-phenylbutanal
-
-
-
-
?
trans-p-chloro-beta-nitrostyrene + isobutanal
3-(4-chlorophenyl)-2,2-dimethyl-4-nitrobutanal
-
-
-
-
?
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate
(3E)-2-oxohex-3-enedioate
-
-
-
-
r
2-hydroxymuconate
2-oxo-3-hexenedioate
-
the conjugated enol, 2-hydroxymuconate is an unusually stable dienol that is reportedly generated in the course of bacterial catabolism of catechol by the enzymes of the meta-fission pathway
the dienol ketonizes chemically in aqueous solution and enzymatically by the action of 4-oxalocrotonate tautomerase to either the beta,gamma-unsaturated ketone or its alpha,beta-conjugated isomer 2-oxo-3-trans-hexenedioate
-
r
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
isomerization of unconjugated 2-oxo acids such as 2-oxo-4-hexenedioate, to its conjugated isomer via dienol intermediate 2-hydroxy-2,4-hexadienedioate, i.e. 2-hydroxymuconate
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
ketonization process via dienol intermediate 2-hydroxymuconate and with Pro1 as a general base, one-proton transfer mechanism
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
via dienol intermediate 2-hydroxymuconate
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
via the intermediate 2-hydroxy-2,4-hexadienedioate, i.e. 2-hydroxymuconate
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
the enzyme converts the unconjugated enone to the conjugated enone via a dienolic intermediate 2-hydroxymuconate, Pro1 serves as the general base, and both Arg11 and Arg39 function in substrate binding and catalysis in an otherwise hydrophobic active site. Anticooperativity during catalysis
-
-
r
additional information
?
-
-
ketonization of two monoacid substrates, 2-hydroxy-2,4-pentadienoate and phenylenolpyruvate, produces a mixture of stereoisomers, 2-keto-3-[2H]-4-pentenoate and 3-[2H]-phenylpyruvate, where the (3R)-isomers predominate
-
-
?
additional information
?
-
-
the achiral substrate 2-hydroxymuconate is processed with equal efficiency by either the D- or the L-enzyme, stereochemical analysis of the D-4OT-catalyzed reaction, overviewn
-
-
?
additional information
?
-
-
no activity with (E)-2-cyclohexyl-1-nitroethene
-
-
?
additional information
?
-
-
the enzyme also has a low level trans-3-chloroacrylic acid dehalogenase activity
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate
(3E)-2-oxohex-3-enedioate
-
-
-
r
2-oxo-4(E)-hexenedioate
2-oxo-3(E)-hexenedioate
-
1,3-allylic isomerization
-
-
?
(2Z,4E)-2-hydroxyhexa-2,4-dienedioate
(3E)-2-oxohex-3-enedioate
-
-
-
-
r
2-hydroxymuconate
2-oxo-3-hexenedioate
-
the conjugated enol, 2-hydroxymuconate is an unusually stable dienol that is reportedly generated in the course of bacterial catabolism of catechol by the enzymes of the meta-fission pathway
the dienol ketonizes chemically in aqueous solution and enzymatically by the action of 4-oxalocrotonate tautomerase to either the beta,gamma-unsaturated ketone or its alpha,beta-conjugated isomer 2-oxo-3-trans-hexenedioate
-
r
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
isomerization of unconjugated 2-oxo acids such as 2-oxo-4-hexenedioate, to its conjugated isomer via dienol intermediate 2-hydroxy-2,4-hexadienedioate, i.e. 2-hydroxymuconate
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
ketonization process via dienol intermediate 2-hydroxymuconate and with Pro1 as a general base, one-proton transfer mechanism
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
via dienol intermediate 2-hydroxymuconate
-
-
?
2-oxo-4-hexenedioate
2-oxo-3-hexenedioate
-
via the intermediate 2-hydroxy-2,4-hexadienedioate, i.e. 2-hydroxymuconate
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2E)-fluoromuconate
-
a dicarboxylate competitive inhibitor, shows substoichiometric binding to 3 +/- 1 sites per hexamer
additional information
-
although 4-OT exhibits no structural asymmetry either by X-ray or by NMR, inactivation by two affinity labels shows half-site stoichiometry
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.334
(2E)-2-hydroxy-3-phenylprop-2-enoate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.18
2-hydroxymuconate
-
mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.44
2-hydroxymuconate
-
mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.512
2-hydroxymuconate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.78
2-hydroxymuconate
-
mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
1.05
2-hydroxymuconate
-
mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
additional information
additional information
-
steady-state kinetics, overview
-
additional information
additional information
-
4-OT exhibits hyperbolic kinetics and shows anticooperativity during catalysis
-
additional information
additional information
-
kinetic modeling, overview
-
additional information
additional information
-
steady-state kinetics of wild-type and mutant 4-OTs for trans-3-chloroacrylic acid dehalogenase activity, pH 8.2, 23°C, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
16
(2E)-2-hydroxy-3-phenylprop-2-enoate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.2
2-hydroxymuconate
-
mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
1.5
2-hydroxymuconate
-
mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
14
2-hydroxymuconate
-
mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
360
2-hydroxymuconate
-
mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
1850
2-hydroxymuconate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
48
(2E)-2-hydroxy-3-phenylprop-2-enoate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
0.45
2-hydroxymuconate
-
mutant enzyme R39A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
8.3
2-hydroxymuconate
-
mutant enzyme P1A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
13
2-hydroxymuconate
-
mutant enzyme R11A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
460
2-hydroxymuconate
-
mutant enzyme R61A, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
3600
2-hydroxymuconate
-
wild type enzyme, in 10 mM potassium phosphate buffer, pH 7.3, at 22°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.045
(2E)-fluoromuconate
-
pH and temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.3
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
-
introduction of polar residues into the active site produces significant decreases in kcat and Km
metabolism
physiological function
additional information
evolution
-
4-oxalocrotonate tautomerase is a member of the tautomerase superfamily
evolution
-
mechanism and the evolution of 4-oxalocrotonate tautomerase, and 5-(carboxymethyl)-2-hydroxymuconate isomerase, EC 5.3.3.10, and their respective pathways, overview
metabolism
-
4-oxalocrotonate tautomerase is an essential enzyme in the degradative metabolism pathway occurring in the Krebs cycle
metabolism
-
4-oxalocrotonate tautomerase is part of a set of inducible enzymes that converts aromatic hydrocarbons to intermediates in the Krebs cycle
metabolism
-
in the catechol meta-fission pathway elaborated by Pseudomonas putida mt-2 ketonization of 2-hydroxymuconate by 4-oxalocrotonate tautomerase generates the alpha,beta-unsaturated ketone 2-oxo-3-(E)-hexenedioate, which undergoes decarboxylation and further processing to intermediates in the Krebs cycle
metabolism
-
the enzyme is part of a degradative pathway that converts various aromatic hydrocarbons to intermediates in the Krebs cycle
physiological function
-
4-OT catalyzes the ketonization process of 2-oxo-4-hexenedioate to its conjugated isomer, 2-oxo-3-hexadienedioate, through the dienol intermediate 2-hydroxymuconate. This proton transfer process is an essential part of degradative metabolism pathway to convert various aromatic hydrocarbons into their corresponding intermediates in the Krebs cycle
physiological function
-
4-oxalocrotonate tautomerase is an extremely efficient catalyst apparently processing either isomer near the diffusion control limit of a small molecule and an enzyme
additional information
-
immediate nonenzymatic conversion of 2-oxo-3-hexenedioate to 2-hydroxy-3-trans-hexenedioate with NaBH4. Rate constants for the nonenzymatic phosphate-catalyzed ketonization of 2-hydroxymuconate, overview
additional information
-
structure-function relationship and kinetic analysis, detailed overview
additional information
-
structure-function relationship, spectroscopic NMR analysis, detailed overview. Three arginine residues, Arg11, Arg39, and Arg61, are localized in the active site of 4-oxalocrotonate tautomerase. Importance of Arg11 in properly orienting the dicarboxylate substrate by interacting with the charged 6-carboxylate group. Arg39 interacts with the 1-carboxylate and the 2-keto group of the substrate to promote carbonyl polarization and catalysis, while Pro-1 transfers protons from C-3 to C-5. Arg61 does not play a significant role in either substrate binding or catalysis
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexamer
additional information
hexamer
-
4-OT has an amino-terminal proline and a beta-alpha-beta fold, three structural models of 4-OT, overview
hexamer
-
the noncovalent structures of each 4OT enantiomer are similar, the predominant form of the enzyme is a hexameric complex of identical subunits
additional information
-
structure determinatin and analysis by NMR of wild-type and mutant L8R 4-OTs, overview
additional information
-
structure-function relationship, NMR analysis, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
mutant enzyme M45Y/F50A, sitting drop vapor diffusion method, using 0.2 M sodium formate, 0.1 M bis-tris propane pH 8.5 and 20% PEG 3350 (w/v)
sitting drop vapor diffusion method, using 100 mM magnesium acetate/100 mM sodium acetate, 5%-21% (w/v) PEG 8000, pH 4.5, and 100 mM calcium acetate/100 mM sodium acetate, 1%-13% (w/v) PEG 4000, pH 4.5
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A33D
the mutant shows highest enantioselectivity for the Michael-type addition of acetaldehyde to trans-beta nitrostyrene producing 4-nitro-3-phenylbutanal, compared to the wild type enzyme
A33E
the mutation significantly (4fold) improves the activity of the enzyme for the Michael-type addition of acetaldehyde to trans-beta nitrostyrene, compared to the wild type enzyme
H6M
the mutant has about 3fold increased specific activity compared with that of wild type enzyme
H6M/A33E/F50V
the mutation strongly enhances the activity for the Michael-type addition of butanal to trans-beta nitrostyrene, compared to the wild type enzyme
M45Y/F50A
the mutant enzyme displays low Michael-type addition activity compared to the wild type enzyme
R39E
the mutant shows highest enantioselectivity for the Michael-type addition of butanal to trans-beta nitrostyrene, producing 2-ethyl-4-nitro-3-phenylbutanal, compared to the wild type enzyme
alphaI52E
-
site-directed mutagenesis, active site mutant,the mutant shows improved trans-3-chloroacrylic acid dehalogenase activity with a 36fold increase in kcat/Km, largely due to a 110fold decrease in Km, and diminished 4-oxalocrotonate tautomerase activity. The negatively charged group may hinder the formation of the enolate intermediate and may contribute to a decrease in kcat
alphaL8R
-
site-directed mutagenesis, active site mutant, the mutant shows improved trans-3-chloroacrylic acid dehalogenase activity with a 50fold increase in kcat/Km, primarily from an 8.8fold increase in kcat, and diminished 4-oxalocrotonate tautomerase activity with a 5fold decrease in kcat/Km. The increased CaaD activity of L8R-4-OT does not substantially diminish the original 4-OT activity
alphaL8R/I52E
-
site-directed mutagenesis, active site mutant, the mutant shows improved trans-3-chloroacrylic acid dehalogenase activity with a 32fold increase in kcat/Km, largely due to a 23fold decrease in Km, and diminished 4-oxalocrotonate tautomerase activity with a 1700fold decrease in kcat/Km
P1A
-
the mutation results in 430fold decreases in kcat/Km compared to the wild type enzyme
R11A
R39A
R39Q
R61A
R11A
-
site-directed mutagenesis, no kinetic effects of the R11A mutation, the stereoselectivity of the R11A-catalyzed protonation at C-5 of the dicarboxylate substrate decreases, while the stereoselectivity of protonation at C-3 of the monocarboxylate substrate increases in comparison with wild-type 4-OT
R11A
-
the mutation results in 280fold decreases in kcat/Km compared to the wild type enzyme
R39A
-
site-directed mutagenesis, with 2-hydroxymuconate the R39A mutant shows decreased kcat by 125fold and increased Km by 1.5fold
R39A
-
the mutation results in 8000fold decreases in kcat/Km compared to the wild type enzyme
R39Q
-
site-directed mutagenesis, with 2-hydroxymuconate the R39Q mutant shows decreased kcat by 389fold and increased Km by 2.6fold, only the tight binding sites function catalytically in the R39Q mutant, structural changes in the R39Q mutant were mainly in the beta-hairpin from residues 50 to 57 which covers the active site
R61A
-
site-directed mutagenesis, Arg61 mutation does not affect either substrate binding or catalysis
R61A
-
the mutation results in 8fold decreases in kcat/Km compared to the wild type enzyme
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
phenyl-Sepharose column chromatography and DEAE-Sepharose column chromatography
-
recombinant enzyme from Escherichia coli by ultracentrifugation, two different steps of anion exchange chromatography, hydroxylapatite and hydrophobic interaction chromatography, and gel filtration with alternating steps of ultrafiltration
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21-Gold(DE3) to over 95% purity
-
recombinant wild-type and mutants from Escherichia coli strain BL21(DE3)pLysS to homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
-
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21-Gold(DE3)
-
overexpression of wild-type and mutants in Escherichia coli strain BL21(DE3)pLysS, subcloning in Escherichia coli strain JM109
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Harris, T.K.; Czerwinski, R.M.; Johnson, W.H.; Legler, P.M.; Abeygunawardana, C.; Massiah, M.A.; Stivers, J.T.; Whitman, C.P.; Mildvan, A.S.
Kinetic, stereochemical, and structural effects of mutations of the active site arginine residues in 4-oxalocrotonate tautomerase
Biochemistry
38
12343-12357
1999
Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
Azurmendi, H.F.; Miller, S.G.; Whitman, C.P.; Mildvan, A.S.
Half-of-the-sites binding of reactive intermediates and their analogues to 4-oxalocrotonate tautomerase and induced structural asymmetry of the enzyme
Biochemistry
44
7725-7737
2005
Pseudomonas putida
Poelarends, G.J.; Almrud, J.J.; Serrano, H.; Darty, J.E.; Johnson, W.H.; Hackert, M.L.; Whitman, C.P.
Evolution of enzymatic activity in the tautomerase superfamily: mechanistic and structural consequences of the L8R mutation in 4-oxalocrotonate tautomerase
Biochemistry
45
7700-7708
2006
Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
Wang, S.; Johnson Jr., W.; Czerwinski, R.; Stamps, S.; Whitman, C.
Kinetic and stereochemical analysis of YwhB, a 4-oxalocrotonate tautomerase homologue in Bacillus subtilis: Mechanistic implications for the YwhB- and 4-oxalocrotonate tautomerase-catalyzed reactions
Biochemistry
46
11919-11929
2007
Bacillus subtilis, Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
Whitman, C.; Aird, B.; Gillespie, W.; Stolowich, N.
Chemical and enzymatic ketonization of 2-hydroxymuconate, a conjugated enol
J. Am. Chem. Soc.
113
3154-3162
1991
Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
-
Whitman, C.; Hajipour, G.; Watson, R.; Johnson Jr., W.; Bembenek, M.; Stolowich, N.
Stereospecific ketonization of 2-hydroxymuconate by 4-oxalocrotonate tautomerase and 5-(carboxymethyl)-2-hydroxymuconate isomerase
J. Am. Chem. Soc.
114
10104-10110
1992
Escherichia coli, Escherichia coli C, Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
-
Fitzgerald, M.; Chernushevich, I.; Standing, K.; Kent, S.; Whitman, C.
Total chemical synthesis and catalytic properties of the enzyme enantiomers L- and D-4-oxalocrotonate tautomerase
J. Am. Chem. Soc.
117
11075-11080
1995
Pseudomonas putida, Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182
-
Wu, P.; Cisneros, G.A.; Hu, H.; Chaudret, R.; Hu, X.; Yang, W.
Catalytic mechanism of 4-oxalocrotonate tautomerase: significances of protein-protein interactions on proton transfer pathways
J. Phys. Chem. B
116
6889-6897
2012
Pseudomonas putida
Burks, E.A.; Yan, W.; Johnson, W.H.; Li, W.; Schroeder, G.K.; Min, C.; Gerratana, B.; Zhang, Y.; Whitman, C.P.
Kinetic, crystallographic, and mechanistic characterization of TomN: elucidation of a function for a 4-oxalocrotonate tautomerase homologue in the tomaymycin biosynthetic pathway
Biochemistry
50
7600-7611
2011
Pseudomonas putida
Narancic, T.; Radivojevic, J.; Jovanovic, P.; Francuski, D.; Bigovic, M.; Maslak, V.; Savic, V.; Vasiljevic, B.; O'Connor, K.E.; Nikodinovic-Runic, J.
Highly efficient Michael-type addition of acetaldehyde to beta-nitrostyrenes by whole resting cells of Escherichia coli expressing 4-oxalocrotonate tautomerase
Biores. Technol.
142
462-468
2013
Pseudomonas putida
Huddleston, J.P.; Burks, E.A.; Whitman, C.P.
Identification and characterization of new family members in the tautomerase superfamily analysis and implications
Arch. Biochem. Biophys.
564
189-196
2014
Pseudomonas putida
Stack, T.M.M.; Li, W.; Johnson, W.H.; Zhang, Y.J.; Whitman, C.P.
Inactivation of 4-oxalocrotonate tautomerase by 5-halo-2-hydroxy-2,4-pentadienoates
Biochemistry
57
1012-1021
2018
Leptothrix cholodnii, Pseudomonas putida (Q01468), Leptothrix cholodnii SP-6
Djokic, L.; Spasic, J.; Jeremic, S.; Vasiljevic, B.; Prodanovic, O.; Prodanovic, R.; Nikodinovic-Runic, J.
Immobilization of Escherichia coli cells expressing 4-oxalocrotonate tautomerase for improved biotransformation of beta-nitrostyrene
Bioprocess Biosyst. Eng.
38
2389-2395
2015
Pseudomonas putida
Poddar, H.; Rahimi, M.; Geertsema, E.M.; Thunnissen, A.M.; Poelarends, G.J.
Evidence for the formation of an enamine species during aldol and Michael-type addition reactions promiscuously catalyzed by 4-oxalocrotonate tautomerase
ChemBioChem
16
738-741
2015
Pseudomonas putida (Q01468)
Baas, B.J.; Zandvoort, E.; Wasiel, A.A.; Poelarends, G.J.
Demethionylation of Pro-1 variants of 4-oxalocrotonate tautomerase in Escherichia coli by co-expression with an engineered methionine aminopeptidase
FEBS Open Bio
4
651-658
2014
Pseudomonas putida
Lazic, J.; Spasic, J.; Francuski, D.; Tokic-Vujosevic, Z.; Nikodinovic-Runic, J.; Maslak, V.; Djokic, L.
Importance of N-terminal proline for the promiscuous activity of 4-oxalocrotonate tautomerase (4-OT)
J. Serb. Chem. Soc.
81
871-881
2016
Pseudomonas putida
-
van der Meer, J.Y.; Poddar, H.; Baas, B.J.; Miao, Y.; Rahimi, M.; Kunzendorf, A.; van Merkerk, R.; Tepper, P.G.; Geertsema, E.M.; Thunnissen, A.M.; Quax, W.J.; Poelarends, G.J.
Using mutability landscapes of a promiscuous tautomerase to guide the engineering of enantioselective Michaelases
Nat. Commun.
7
10911
2016
Pseudomonas putida (Q01468)
Radivojevic, J.; Minovska, G.; Senerovic, L.; OConnor, K.; Jovanovic, P.; Savic, V.; Tokic-Vujosevic, Z.; Nikodinovic-Runic, J.; Maslak, V.
Synthesis of gamma-nitroaldehydes containing quaternary carbon in the alpha-position using a 4-oxalocrotonate tautomerase whole-cell biocatalyst
RSC Adv.
4
60502-60510
2014
Pseudomonas putida
-
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