Information on EC 4.1.3.17 - 4-hydroxy-4-methyl-2-oxoglutarate aldolase

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

EC NUMBER
COMMENTARY hide
4.1.3.17
-
RECOMMENDED NAME
GeneOntology No.
4-hydroxy-4-methyl-2-oxoglutarate aldolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-hydroxy-4-oxobutane-1,2,4-tricarboxylate = oxaloacetate + pyruvate
show the reaction diagram
(2)
-
-
-
4-hydroxy-4-methyl-2-oxoglutarate = 2 pyruvate
show the reaction diagram
(1)
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-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
of an oxo-acid, C-C bond cleavage
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-hydroxy-4-methyl-L-glutamate biosynthesis
-
-
Benzoate degradation
-
-
C5-Branched dibasic acid metabolism
-
-
gallate degradation
-
-
gallate degradation I
-
-
gallate degradation II
-
-
methylgallate degradation
-
-
Microbial metabolism in diverse environments
-
-
protocatechuate degradation I (meta-cleavage pathway)
-
-
SYSTEMATIC NAME
IUBMB Comments
4-hydroxy-4-methyl-2-oxoglutarate pyruvate-lyase (pyruvate-forming)
Requires a divalent metal ion [3]. This enzyme participates in the degradation of protocatechuate (via the meta-cleavage pathway), phthalate, syringate and gallate [1-3]. The enzyme from Pseudomonas ochraceae can also cleave 4-hydroxy-2-oxoglutarate to glyoxylate and pyruvate, and also catalyses the reaction of EC 4.1.1.3 (oxaloacetate decarboxylase) [3].
CAS REGISTRY NUMBER
COMMENTARY hide
37290-65-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
development of a selection strategy for the evolution of the substrate specificity of KDPG aldolase based on a pyruvate kinase-deficient strain of Escherichia coli
-
-
Manually annotated by BRENDA team
strain NGJ1
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
-
4-hydroxy-4-methyl-2-oxoglutarate/4-carboxy-4-hydroxy-2-oxoadipate aldolases are class II pyruvate aldolases from the meta cleavage pathways of protocatechuate and gallate
additional information
-
the Pseudomonas putida F1 HMG/CHA aldolase has a D-X20-R-D motif, active site structure, structure-function relationship, overview. The Glu199 residue in HMG/CHA aldolase positions one water molecule and in concert with the Asp102 residue positions a second water molecule that coordinate with the bound magnesium ion
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-hydroxy-4-oxopentanedioic acid
?
show the reaction diagram
-
-
-
-
?
4-carboxy-4-hydroxy-2-oxoadipate
pyruvate + oxaloacetate
show the reaction diagram
4-Hydroxy-2-oxoglutarate
Pyruvate + glyoxylate
show the reaction diagram
4-hydroxy-2-oxopentanoate
?
show the reaction diagram
-
-
-
-
?
4-hydroxy-4-methyl-2-oxoglutarate
2 pyruvate
show the reaction diagram
4-hydroxy-4-methyl-2-oxoglutarate
pyruvate
show the reaction diagram
-
involved in biosynthesis of gamma-substituted glutamic acid, participates in the alpha-keto acid pathway for degradation of the meta-fission product of protocatechuate, in Pseudomonas ochraceae grown on phthalate the physiolological substrate is not hydroxymethyl glutamic acid but CHA, L-4-carboxy-4-hydroxy-2-oxoadipate, which is cleaved to pyruvate and oxaloacetate
-
r
4-hydroxy-4-methyl-2-oxoglutarate
pyruvate + pyruvate
show the reaction diagram
D-4-carboxy-4-hydroxy-2-oxoadipate
pyruvate + oxaloacetate
show the reaction diagram
DL-4-carboxy-4-hydroxy-2-oxoadipate
pyruvate + oxaloacetate
show the reaction diagram
DL-4-hydroxy-2-oxoglutarate
pyruvate + glyoxylate
show the reaction diagram
-
-
-
?
DL-4-hydroxy-4-methyl-2-oxoglutarate
pyruvate
show the reaction diagram
l-4-carboxy-4-hydroxy-2-oxoadipate
pyruvate + oxaloacetate
show the reaction diagram
oxaloacetate
?
show the reaction diagram
pyruvate + glyoxylate
4-hydroxy-2-oxoglutarate
show the reaction diagram
-
-
-
-
pyruvate + pyruvate
4-hydroxy-4-methyl-2-oxoglutarate
show the reaction diagram
additional information
?
-
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-4-methyl-2-oxoglutarate
2 pyruvate
show the reaction diagram
-
-
-
-
?
4-hydroxy-4-methyl-2-oxoglutarate
pyruvate
show the reaction diagram
-
involved in biosynthesis of gamma-substituted glutamic acid, participates in the alpha-keto acid pathway for degradation of the meta-fission product of protocatechuate, in Pseudomonas ochraceae grown on phthalate the physiolological substrate is not hydroxymethyl glutamic acid but CHA, L-4-carboxy-4-hydroxy-2-oxoadipate, which is cleaved to pyruvate and oxaloacetate
-
r
l-4-carboxy-4-hydroxy-2-oxoadipate
pyruvate + oxaloacetate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
absolute requirement for divalent metal ion: Mg2+, Mn2+, Co2+, Zn2+ or Cd2+; Km: 0.0034 mM, reaction with L-4-carboxy-4-hydroxy-2-oxoadipate
Zn2+
-
absolute requirement for divalent metal ion: Mg2+, Mn2+, Co2+, Zn2+ or Cd2+, Km: 0.0018 mM, reaction with L-4-carboxy-4-hydroxy-2-oxoadipate
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoadipate
-
competitive with respect to DL-4-carboxy-4-hydroxy-2-oxoadipate
2-oxobutanoate
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competitive with respect to DL-4-carboxy-4-hydroxy-2-oxoadipate
2-oxoglutarate
4-hydroxy-2-oxobutyrate
-
competitive with respect to DL-4-carboxy-4-hydroxy-2-oxoadipate
ATP
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competitive
citrate
-
competitive
diethyl dicarbonate
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not reversed by hydroxylamine, L-4-carboxy-4-hydroxy-2-oxoadipate strongly protects against inactivation
DL-4-hydroxy-2-oxovalerate
-
competitive with respect to DL-4-carboxy-4-hydroxy-2-oxoadipate
HgCl2
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0.1 mM, 41% inhibition
NEM
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1 mM, 10% inhibition
oxalate
oxaloacetate
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competitive
PCMB
-
1 mM, 48% inhibition
pyruvic aldol lactone
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-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
phosphate
-
activates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.071 - 0.15
2-hydroxy-4-oxopentanedioic acid
0.29
4-carboxy-4-hydroxy-2-oxoadipate
-
-
8.8 - 25
4-hydroxy-2-oxopentanoate
0.022 - 5.5
4-hydroxy-4-methyl-2-oxoglutarate
0.097
D-4-carboxy-4-hydroxy-2-oxoadipate
-
activated by Mg2+
0.044
DL-4-carboxy-4-hydroxy-2-oxoadipate
-
pH 8.0, 25C
0.24
DL-4-hydroxy-2-oxoglutarate
-
pH 8.0, 25C
1.25
DL-4-hydroxy-4-methyl-2-oxoglutarate
-
pH 8.0, 25C
0.0065 - 0.15
L-4-carboxy-4-hydroxy-2-oxoadipate
0.036 - 0.5
oxaloacetate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.058 - 0.25
2-hydroxy-4-oxopentanedioic acid
0.048 - 0.19
4-hydroxy-2-oxopentanoate
0.0276 - 19.3
4-hydroxy-4-methyl-2-oxoglutarate
1.52 - 1.85
oxaloacetate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38 - 3.5
2-hydroxy-4-oxopentanedioic acid
0.0019 - 0.022
4-hydroxy-2-oxopentanoate
74 - 83000
4-hydroxy-4-methyl-2-oxoglutarate
6.2 - 420
oxaloacetate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0017 - 0.0377
oxalate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37.7
-
wild type protein, pH 8.0, 25C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.6
-
L-4-carboxy-4-hydroxy-2-oxoadipate, without phosphate
7.7
-
L-4-carboxy-4-hydroxy-2-oxoadipate, in presence of 0.2 mM phosphate
8.2
-
L-4-carboxy-4-hydroxy-2-oxoadipate, with 3 mM phosphate
8.4
-
DL-4-hydroxy-4-methyl-2-oxoglutarate, in presence of 0.2 mM phosphate
8.6
-
D-4-carboxy-4-hydroxy-2-oxoadipate, with 3 mM phosphate
8.8
-
oxaloacetate, without phosphate
9.4
-
DL-4-hydroxy-2-oxoglutarate, in presence of 0.2 mM oxoglutarate
additional information
-
broad optimum (log KCAT) pH 8 - pH 10.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 10.5
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-
7.5 - 9.5
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pH 7.5: about 30% of maximal activity, pH 9.5: about 65% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
isoelectric focusing
5.7
-
predicted pI
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas putida (strain F1 / ATCC 700007)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24070
-
calculated from amino acid sequence corresponding to gene sequence
150000
-
equilibrium sedimentation
160000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
homohexamer
-
6 * 26000, crystal structure, gel filtration and SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop method, PEG 4000
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
54C, 10 min, 50% loss of activity
33192
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
54
-
pH 7.0, 10 min, 50% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, 20 mM sodium HEPES buffer, pH7.5, stable for 2 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ion exchange chromatography, hydrophobic interaction chromatography, gel filtration
-
wild-type and recombinant enzyme
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA fragment carrying the gene proA from chromosomal DNA cloned and overproduced in Escherichia coli XL1-Blue MRF'
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expressed in Escherichia coli BL21(lambdaDE3)
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phylogenetic analysis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E199A
-
site-directed mutagenesis, the mutant shows reduced activity and reduced affinity for Co2+ compared to the wild-type enzyme
R123A
-
no activity
R123K
-
retains some activity
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