Information on EC 4.1.3.42 - (4S)-4-hydroxy-2-oxoglutarate aldolase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
4.1.3.42
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RECOMMENDED NAME
GeneOntology No.
(4S)-4-hydroxy-2-oxoglutarate aldolase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(4S)-4-hydroxy-2-oxoglutarate = pyruvate + glyoxylate
show the reaction diagram
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-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glyoxylate and dicarboxylate metabolism
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SYSTEMATIC NAME
IUBMB Comments
(4S)-4-hydroxy-2-oxoglutarate glyoxylate-lyase (pyruvate-forming)
The enzyme from the bacterium Escherichia coli is specific for the (S) enantiomer. That enzyme is trifunctional, and also catalyses the reactions of EC 4.1.1.3, oxaloacetate decarboxylase and EC 4.1.2.14, 2-dehydro-3-deoxy-phosphogluconate aldolase. cf. EC 4.1.3.16, 4-hydroxy-2-oxoglutarate aldolase.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
bovine
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-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(4R)-4-hydroxy-2-oxoglutarate
pyruvate + glyoxylate
show the reaction diagram
poor substrate
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-
r
(4S)-4-hydroxy-2-oxobutanoate
formaldehyde
show the reaction diagram
at 8% of the rate with DL-4-hydroxy-2-oxoglutarate
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-
r
(4S)-4-hydroxy-2-oxoglutarate
pyruvate + glyoxylate
show the reaction diagram
2-dehydro-3-deoxy-6-phospho-D-gluconate
?
show the reaction diagram
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-
-
-
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2-Keto-4-hydroxybutyrate
Pyruvate + formaldehyde
show the reaction diagram
4-hydroxy-2-oxoglutarate
?
show the reaction diagram
4-Hydroxy-2-oxoglutarate
Pyruvate + glyoxylate
show the reaction diagram
D,L-4-hydroxy-2-oxoglutarate
pyruvate + glyoxylate + D-4-hydroxy-2-oxoglutarate
show the reaction diagram
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preferential cleavage of the L-isomer of racemic 4-hydroxy-2-oxoglutarate leaving the D-isomer in solution
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r
Glyoxylate + pyruvate
4-Hydroxy-2-ketoglutarate
show the reaction diagram
higher specificity for pyruvate than for glyoxylate
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Oxaloacetate
CO2 + pyruvate
show the reaction diagram
pyruvate + glyoxylate
(4S)-4-hydroxy-2-oxoglutarate
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-hydroxy-2-oxoglutarate
?
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-Ketoglutarate
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4-(Oxyacetyl)phenoxyacetic acic
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inactivates arginine residues, 90% inactivation at 10 mM
4-mercuribenzenesulfonic acid
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1 mM, 56% inhibition
acetaldehyde
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Ki: 11 mM
Bromopyruvate
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85% inhibition at 1 mM, pyruvate or 2-keto-4-hydroxyglutarate protect; substrate analog. Treatment results in a time- and concentration-dependent loss of enzymatic activity. The substrates pyruvate and 2-keto-4-hydroxyglutarate provide more than 90% protection against inactivation by bromopyruvate, no protective effect is seen with glycolaldehyde. 1.1 mol of bromopyruvate is incorporated per enzyme subunit by esterification of residue Glu45
chloride
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41% inhibition at 40 mM
citrate
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Ki: 7.5 mM
CN-
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irreversible, in presence of 4-hydroxy-2-oxoglutarate; irreversible loss of activity in presence of glyoxylate, but not in presence of pyruvate
glyoxylate
Halides
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Hydroxypyruvate
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Ki: 0.0125 mM
iodoacetate
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46% inhibition at 5 mM; 50 mM, almost complete inhibition
N-ethylmaleimide
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10% inhibition at 2.5 mM
NaBH4
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; extensive loss of activity after incubation of enzyme with NaBH4 in presence of pyruvate or glyoxylate
NaBr
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48% inhibition at 20 mM
NaCl
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38% inhibition at 20 mM
NaF
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19% inhibition at 20 mM
NaI
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59% inhibition at 20 mM
p-Mercuriphenylsulfonate
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50% inhibition at 0.5 mM
pyruvate
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competitive inhibition of glyoxylate binding
Sulfhydryl-reacting reagents
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partial
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
25
(4R)-4-hydroxy-2-oxoglutarate
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pH 8.1, 25C
2.3 - 3.5
(4S)-4-hydroxy-2-oxoglutarate
2.5 - 25
(D)-4-hydroxy-alpha-ketoglutarate
2.3
(L)-4-hydroxy-alpha-ketoglutarate
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-
0.039
4-hydroxy-alpha-ketoglutarate
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14.3
glyoxylate
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45
pyruvate
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additional information
additional information
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.5
citrate
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0.0125
Hydroxypyruvate
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0.5
p-Mercuriphenylsulfonate
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.53
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(D)-4-hydroxy-alpha-ketoglutarate
7.9
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(L)-4-hydroxy-alpha-ketoglutarate
45
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pH 8.4, 25C
96.7
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pH 8.1, 25C
additional information
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80fold increase in specific activity after overexpression in pKK223.3
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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2.2 to 2.4 times higher activity is measured in Tris-HCl buffer than in potassium phosphate buffer at the same pH values
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4 - 9.6
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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enzyme assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21169
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3 * 21000, SDS-PAGE, 3 * 21169, calculated
22000
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3 * 22284, calculated, 3 * 22000, SDS-PAGE
22284
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3 * 22284, calculated, 3 * 22000, SDS-PAGE
22286
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3 * 22286, amino acid sequence analysis, identical
24000
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3 * 24000, SDS-PAGE
66860
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amino acid sequence analysis
68000
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PAGE
120000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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trimer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.6
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95% loss of activity after 10 min at 4C, reactivation within 20 min at pH 6-8
5251
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
exposure to phosphoric acid at pH 1.6 for 10 min at 4C causes 95% or greater inactivation. Chloride ion at 50-100 mM or 10 mM 2-oxo-4-hydroxyglutarate markedly decreases both the rate and extent of inactivation; good protection is also afforded by 10 mM pyruvate, glyoxylate, glyoxal, 2-oxoglutarate or 2-oxobutyrate. The inactivation process is characterized by an alteration of secondary and tertiary structure as well as an aggregation to a dimer of the native molecule. Reactivation of enzyme activity to 60-80% of the original level is seen within 20 min at pH 6 to 8
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irreversibly inactivated by CN- in presence of 4-hydroxy-2-oxoglutarate
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, Tris-HCl buffer, pH 7.4, 40-50% loss of activity after 1 month, more labile when frozen
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5C, potassium phosphate buffer, pH 7.4, one week, sometimes several months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
strain K-12
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; overexpressed in pKK223.3 expression vector
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
exposure to phosphoric acid at pH 1.6 for 10 min at 4C causes 95% or greater inactivation. The inactivation process is characterized by an alteration of secondary and tertiary structure as well as an aggregation to a dimer of the native molecule. Reactivation of enzyme activity to 60-80% of the original level is seen within 20 min at pH 6 to 8
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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preparation of (4S)-4-hydroxy-2-oxoglutarate at more than 95% enantiomeric excess by stereospecific synthesis from glyoxylate and pyruvate. Preparation of (4R)-4-hydroxy-2-oxoglutarate at 60% enantiomeric excess by selective cleavage of the (4S)-isomer of racemic 4-hydroxy-2-oxoglutarate leaving the (4R)-isomer in solution