BRENDA - Enzyme Database show
show all sequences of 4.1.3.42

Purification, substrate specificity and binding, beta-decarboxylase activity, and other properties of Escherichia coli 2-keto-4-hydroxyglutarate aldolase

Nishihara, H.; Dekker, E.E.; J. Biol. Chem. 247, 5079-5087 (1972)

Data extracted from this reference:

General Stability
General Stability
Organism
irreversibly inactivated by CN- in presence of 4-hydroxy-2-oxoglutarate
Escherichia coli
Inhibitors
Inhibitors
Commentary
Organism
Structure
4-mercuribenzenesulfonic acid
1 mM, 56% inhibition
Escherichia coli
CN-
irreversible, in presence of 4-hydroxy-2-oxoglutarate; irreversible loss of activity in presence of glyoxylate, but not in presence of pyruvate
Escherichia coli
iodoacetate
46% inhibition at 5 mM; 50 mM, almost complete inhibition
Escherichia coli
additional information
not inhibitory: EDTA, 1,10-phenanthroline, S-hydroxyquinoline, or alpha,alpha'-dipyridyl
Escherichia coli
N-ethylmaleimide
10% inhibition at 2.5 mM
Escherichia coli
NaBH4
; extensive loss of activity after incubation of enzyme with NaBH4 in presence of pyruvate or glyoxylate
Escherichia coli
p-Mercuriphenylsulfonate
50% inhibition at 0.5 mM
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Escherichia coli
2.3
-
(4S)-4-hydroxy-2-oxoglutarate
pH 8.1, 25C
Escherichia coli
2.3
-
(L)-4-hydroxy-alpha-ketoglutarate
-
Escherichia coli
2.5
-
(D)-4-hydroxy-alpha-ketoglutarate
-
Escherichia coli
25
-
(4R)-4-hydroxy-2-oxoglutarate
pH 8.1, 25C
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
additional information
divalent kations are not required. No effect on activity: Mg2+, Mn2+, Zn2+, Ca2+, Ba2+, Co2+, Cu2+, Ni2+, Fe2+, Hg2+, K+, or Fe3+; no requirement
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
62500
-
gel filtration; gel filtration
Escherichia coli
64000
-
sucrose density gradient centrifugation; sucrose density gradient centrifugation
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-hydroxy-2-oxoglutarate
Escherichia coli
possibly involved in regulation of tricarboxylic acid cycle
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Escherichia coli
P0A955
-
-
Purification (Commentary)
Commentary
Organism
;
Escherichia coli
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.53
-
(D)-4-hydroxy-alpha-ketoglutarate
Escherichia coli
7.9
-
(L)-4-hydroxy-alpha-ketoglutarate
Escherichia coli
96.7
-
pH 8.1, 25C
Escherichia coli
Storage Stability
Storage Stability
Organism
4C, Tris-HCl buffer, pH 7.4, 40-50% loss of activity after 1 month, more labile when frozen
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(4R)-4-hydroxy-2-oxoglutarate
poor substrate
5242
Escherichia coli
pyruvate + glyoxylate
-
-
-
r
(4S)-4-hydroxy-2-oxobutanoate
at 8% of the rate with DL-4-hydroxy-2-oxoglutarate
5242
Escherichia coli
formaldehyde
-
-
-
r
(4S)-4-hydroxy-2-oxoglutarate
-
5242
Escherichia coli
pyruvate + glyoxylate
L-4-hydroxy-2-oxoglutarate, pyruvate and glyoxylate all bind at the same active site of the enzyme
-
-
r
2-Keto-4-hydroxybutyrate
-
5242
Escherichia coli
Pyruvate + formaldehyde
-
-
-
-
4-Hydroxy-2-oxoglutarate
-
5242
Escherichia coli
Pyruvate + glyoxylate
-
5242
Escherichia coli
-
4-hydroxy-2-oxoglutarate
possibly involved in regulation of tricarboxylic acid cycle
5242
Escherichia coli
?
-
-
-
-
Glyoxylate + pyruvate
higher specificity for pyruvate than for glyoxylate
5242
Escherichia coli
4-Hydroxy-2-ketoglutarate
-
-
-
-
additional information
highly specific towards L-isomer
5242
Escherichia coli
?
-
-
-
-
additional information
enzyme additionally shows beta-decarboxylase activity towards oxalacetate
5242
Escherichia coli
?
-
-
-
?
Oxaloacetate
better carboxylase than aldolase
5242
Escherichia coli
CO2 + pyruvate
-
5242
Escherichia coli
-
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
enzyme assay at
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
2.2 to 2.4 times higher activity is measured in Tris-HCl buffer than in potassium phosphate buffer at the same pH values
Escherichia coli
8.6
-
;
Escherichia coli
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.5
-
p-Mercuriphenylsulfonate
-
Escherichia coli
General Stability (protein specific)
General Stability
Organism
irreversibly inactivated by CN- in presence of 4-hydroxy-2-oxoglutarate
Escherichia coli
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
4-mercuribenzenesulfonic acid
1 mM, 56% inhibition
Escherichia coli
CN-
irreversible, in presence of 4-hydroxy-2-oxoglutarate
Escherichia coli
CN-
irreversible loss of activity in presence of glyoxylate, but not in presence of pyruvate
Escherichia coli
iodoacetate
46% inhibition at 5 mM
Escherichia coli
iodoacetate
50 mM, almost complete inhibition
Escherichia coli
additional information
not inhibitory: EDTA, 1,10-phenanthroline, S-hydroxyquinoline, or alpha,alpha'-dipyridyl
Escherichia coli
N-ethylmaleimide
10% inhibition at 2.5 mM
Escherichia coli
NaBH4
-
Escherichia coli
NaBH4
extensive loss of activity after incubation of enzyme with NaBH4 in presence of pyruvate or glyoxylate
Escherichia coli
p-Mercuriphenylsulfonate
50% inhibition at 0.5 mM
Escherichia coli
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.5
-
p-Mercuriphenylsulfonate
-
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Escherichia coli
2.3
-
(4S)-4-hydroxy-2-oxoglutarate
pH 8.1, 25C
Escherichia coli
2.3
-
(L)-4-hydroxy-alpha-ketoglutarate
-
Escherichia coli
2.5
-
(D)-4-hydroxy-alpha-ketoglutarate
-
Escherichia coli
25
-
(4R)-4-hydroxy-2-oxoglutarate
pH 8.1, 25C
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
additional information
divalent kations are not required. No effect on activity: Mg2+, Mn2+, Zn2+, Ca2+, Ba2+, Co2+, Cu2+, Ni2+, Fe2+, Hg2+, K+, or Fe3+
Escherichia coli
additional information
no requirement
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
62500
-
gel filtration
Escherichia coli
64000
-
sucrose density gradient centrifugation
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4-hydroxy-2-oxoglutarate
Escherichia coli
possibly involved in regulation of tricarboxylic acid cycle
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.53
-
(D)-4-hydroxy-alpha-ketoglutarate
Escherichia coli
7.9
-
(L)-4-hydroxy-alpha-ketoglutarate
Escherichia coli
96.7
-
pH 8.1, 25C
Escherichia coli
Storage Stability (protein specific)
Storage Stability
Organism
4C, Tris-HCl buffer, pH 7.4, 40-50% loss of activity after 1 month, more labile when frozen
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(4R)-4-hydroxy-2-oxoglutarate
poor substrate
5242
Escherichia coli
pyruvate + glyoxylate
-
-
-
r
(4S)-4-hydroxy-2-oxobutanoate
at 8% of the rate with DL-4-hydroxy-2-oxoglutarate
5242
Escherichia coli
formaldehyde
-
-
-
r
(4S)-4-hydroxy-2-oxoglutarate
-
5242
Escherichia coli
pyruvate + glyoxylate
L-4-hydroxy-2-oxoglutarate, pyruvate and glyoxylate all bind at the same active site of the enzyme
-
-
r
2-Keto-4-hydroxybutyrate
-
5242
Escherichia coli
Pyruvate + formaldehyde
-
-
-
-
4-Hydroxy-2-oxoglutarate
-
5242
Escherichia coli
Pyruvate + glyoxylate
-
5242
Escherichia coli
-
4-hydroxy-2-oxoglutarate
possibly involved in regulation of tricarboxylic acid cycle
5242
Escherichia coli
?
-
-
-
-
Glyoxylate + pyruvate
higher specificity for pyruvate than for glyoxylate
5242
Escherichia coli
4-Hydroxy-2-ketoglutarate
-
-
-
-
additional information
highly specific towards L-isomer
5242
Escherichia coli
?
-
-
-
-
additional information
enzyme additionally shows beta-decarboxylase activity towards oxalacetate
5242
Escherichia coli
?
-
-
-
?
Oxaloacetate
better carboxylase than aldolase
5242
Escherichia coli
CO2 + pyruvate
-
5242
Escherichia coli
-
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
enzyme assay at
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
2.2 to 2.4 times higher activity is measured in Tris-HCl buffer than in potassium phosphate buffer at the same pH values
Escherichia coli
8.6
-
-
Escherichia coli
Other publictions for EC 4.1.3.42
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)
5256
Cayrol
Recovery of respiration follow ...
Escherichia coli
Proc. Natl. Acad. Sci. USA
92
11806-11809
1995
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4984
Taha
Purification and characterizat ...
Azotobacter vinelandii
Biochem. Biophys. Res. Commun.
200
459-466
1994
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1
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5258
Patil
Cloning, nucleotide sequence, ...
Escherichia coli
J. Bacteriol.
174
102-107
1992
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1
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1
3
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2
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1
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1
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722866
Floyd
-
A simple strategy for obtainin ...
Escherichia coli
J. Chem. Soc. Perkin Trans. I
1992
1085-1086
1992
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1
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1
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3
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1
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3
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5251
Vlahos
Active-site residues of 2-keto ...
Escherichia coli
J. Biol. Chem.
265
20384-20389
1990
-
-
-
-
-
-
1
1
-
1
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2
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1
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2
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1
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2
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1
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2
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1
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5254
Vlahos
The complete amino acid sequen ...
Escherichia coli
J. Biol. Chem.
263
11683-11691
1988
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1
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2
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722583
Vlahos
Amino acid sequence of the pyr ...
Escherichia coli
J. Biol. Chem.
261
11049-11055
1986
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1
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721748
Winter
Dimerization occurs during the ...
Escherichia coli
Biochim. Biophys. Acta
749
52-61
1983
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5249
Wang
Physical and chemical evidence ...
Escherichia coli
J. Biol. Chem.
256
1793-1800
1981
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5250
Grady
Steady-state kinetics and inhi ...
Escherichia coli
Biochemistry
20
2497-2502
1981
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12
3
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1
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1
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12
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1
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5244
Dekker
-
2-keto-4-Hydroxyglutarate aldo ...
Escherichia coli
Methods Enzymol.
42C
285-290
1978
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2
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1
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1
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5248
Dekker
-
Aldol-type reactions and 2-ket ...
Bos taurus, Escherichia coli
Bioorg. Chem.
1
59-77
1977
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2
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2
3
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2
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1
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12
2
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2
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2
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3
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1
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1
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12
2
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2
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5245
Meloche
Enzyme stereoselectivity: the ...
Escherichia coli
Biochem. Biophys. Res. Commun.
65
1033-1039
1975
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4971
Wood
-
2-Keto-3-deoxy-6-phosphoglucon ...
Bacteria
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
281-302
1972
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5242
Nishihara
Purification, substrate specif ...
Escherichia coli
J. Biol. Chem.
247
5079-5087
1972
-
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1
7
5
-
1
2
1
-
2
-
-
1
-
-
-
3
1
10
-
1
-
-
-
2
-
-
-
1
-
-
-
-
-
-
-
-
1
-
10
1
5
-
2
4
1
-
-
-
2
-
-
3
1
10
-
1
-
-
-
3
-
-
-
-
-
-
-
-
-