Information on EC 4.2.1.130 - D-lactate dehydratase

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

EC NUMBER
COMMENTARY hide
4.2.1.130
-
RECOMMENDED NAME
GeneOntology No.
D-lactate dehydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-lactate = 2-oxopropanal + H2O
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
methylglyoxal degradation
-
-
Pyruvate metabolism
-
-
Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
(R)-lactate hydro-lyase
The enzyme, described from the fungi Candida albicans and Schizosaccharomyces pombe, converts 2-oxopropanal to (R)-lactate in a single glutathione (GSH)-independent step. The other known route for this conversion is the two-step GSH-dependent pathway catalysed by EC 4.4.1.5 (lactoylglutathione lyase) and EC 3.1.2.6 (hydroxyacylglutathione hydrolase).
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
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glyoxalase activity is completely abolished in the gene hchA-deficient strain. Stationary-phase Escherichia coli cells become more susceptible to methylgloxal when gene hchA is deleted, which can be complemented by an expression of plasmid-encoded hch. Accumulation of intracellular methylglyoxal in hchA-deficient strains; the stationary-phase Escherichia coli cells becomes more susceptible to methylglyoxal when hchA is deleted
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-lactate
2-oxopropanal + H2O
show the reaction diagram
2-oxoethanal + H2O
acetate
show the reaction diagram
i.e. glyoxal
-
-
?
2-oxopropanal + H2O
(R)-lactate
show the reaction diagram
acetate
2-oxoethanal + H2O
show the reaction diagram
glyoxal + H2O
acetate
show the reaction diagram
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
phenylglyoxal + H2O
?
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
(R)-lactate
2-oxopropanal + H2O
show the reaction diagram
2-oxopropanal + H2O
(R)-lactate
show the reaction diagram
acetate
2-oxoethanal + H2O
show the reaction diagram
glyoxal + H2O
acetate
show the reaction diagram
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
-
activates; enhances activity
Mg2+
-
activates slightly
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,4-Dinitrophenylhydrazine
inactivation; inactivation
4-hydroxymercuribenzoate
-
inactivates the enzyme, reversible by dithiothreitol and other thiol-group-containing compounds
5,5'-dithiobis-(2-nitrobenzoate)
-
0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
Borate
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boric acid/borate buffer is moderately inhibitory
Cu2+
-
; maximal inhibition below 0.025 mM
EDTA
-
an extensive dialysis of Hsp31 with 10 mM EDTA does not significantly decrease the glyoxalase III activity of more than 30%
glyceraldehyde
about 60% inhibition at 10 mM
glycolaldehyde
about 60% inhibition at 10 mM
glyoxylate
determmination of an enzyme crystal structure with the inhibitor bound to the active Cys residue of the enzyme as a hemithioacetal, detailed binding structure analysis, overview
hydrogen peroxide
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sensitivity of glyoxalase III is special and might relate to the thiol group that is essential for its activity and possibly to the binding of iron adjacent to the active site thiol
N-ethylmaleimide
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0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
Ni2+
-
inhibits slightly
p-hydroxymercuribenzoate
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0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
superoxide
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sensitivity of glyoxalase III is special and might relate to the thiol group that is essential for its activity and possibly to the binding of iron adjacent to the active site thiol
Zn2+
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; maximal inhibition above 0.3 mM, 10% inhibition at 0.025 mM, more than 50% inhibition at 0.1 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bovine serum albumin
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
156.9
2-oxopropanal
-
pH 7.5, 37°C
0.19 - 58
methylglyoxal
13
Phenylglyoxal
pH and temperature not specified in the publication
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15 - 0.42
2-oxoethanal
-
0.08 - 1.43
2-oxopropanal
0.095 - 2.61
methylglyoxal
0.0272
Phenylglyoxal
pH and temperature not specified in the publication
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 1.8
methylglyoxal
0.0021
Phenylglyoxal
pH and temperature not specified in the publication
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.14
glyoxylate
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0055
substrate methylglyoxal, pH 6.8, 37°C
0.0062
substrate methylglyoxal, pH 6.8, 37°C
0.013
substrate methylglyoxal, pH 6.8, 37°C
0.015
substrate methylglyoxal, pH 6.8, 37°C
0.033 - 0.054
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crude enzyme extracts, pH 8.0, 37°C
0.25
substrate glyoxal, pH 6.8, 37°C
0.254
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pH 7.5, temperature not specified in the publication; purified native enzyme, pH 7.5, 37°C
0.31
substrate glyoxal, pH 6.8, 37°C
37
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pH 8.0, 37°C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8
assay at; assay at; assay at; assay at; assay at
7.5
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assay at
8
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assay at; glyoxalase III is active over a wide range of pH with no sharp pH optimum
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 10
-
activity range, profile overview
5 - 10
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glyoxalase III is active over a wide range of pH with no sharp pH optimum, sharp decrease in reaction rate occurs below pH 5.0, 20.25% of maximal activity at pH 10.0; pH 5.0: sharp decrease in reaction rate below, pH 10.0: activity 20-25% lower than the rate at physiological pH
5 - 9
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pH 5.0: about 45% of maximal activity, pH 9.0: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
assay at; assay at
35 - 40
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-
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 50
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activity range, profile overview
25 - 50
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25°C: about 55% of maximal activity, 50°C: about 75% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
glyoxalase III activity is present in all three media in almost equal amounts ranging from 0.033 to 0.054 unit/mg of protein
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Candida albicans (strain SC5314 / ATCC MYA-2876)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
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2 * 40000, SDS-PAGE
80000
gel filtration, recombinant enzyme
82000
-
gel filtration; gel filtration
84000
gel filtration, recombinant enzyme
126000
gel filtration, recombinant enzyme; gel filtration, recombinant enzyme
132000
gel filtration, recombinant enzyme
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
enzyme structure comparisons, overview
trimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme DJ-1d covalently bound to glyoxylate, X-ray diffraction structure determinatin and analysis at 1.60 A resolution, PDB ID 4OGF, and analysis of structure PDB ID 1P5F
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
DTT and either sucrose or mannitol are required for stability
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15°C, 1 mM DTT and 10% sucrose, almost completely stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; native enzyme 27.6fold by ammonium sulfate fractionation, heat treatment, gel filtration chromatography, anion exchange chromatography, and hydrophobic interaction chromatography on n-butyl sepharose resin
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; native enzyme 725fold by ammonium sulfate fractionation, heat treatment, gel filtration, anion exchange chromatography, and affinity chromatography on a 4-hydroxymercuribenzoate-bound resin
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recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression of C-terminally GFP-tagged SpDJ-1, expression of wild-type and mutant SpDj-1 proteins in Escherichia coli strain BL21(DE3), detailed phylogenetic analysis of GLOIII, DJ-1 and HSP31 proteins, overview; gene hsp3101 encoding a Hsp31 protein, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis, recombinant overexpression of Schizosaccharomyces pombe hsp3101 in Schizosaccharomyces pombe wild-type cells or GLO1 deletion cells; gene hsp3101, expression of C-terminally GFP-tagged HSP31 proteins, expression of HSP3101 protein in Escherichia coli strain BL21(DE3), detailed phylogenetic analysis of GLOIII, DJ-1 and HSP31 proteins, overview; gene hsp3102 encoding a Hsp31 protein, sequence comparisons, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant overexpression of Schizosaccharomyces pombe hsp3101 in Schizosaccharomyces pombe wild-type cells or GLO1 deletion cells; gene hsp3106 encoding DJ-1 protein, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis
gene DJ1A, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of HIs-tagged enzyme in Escherichia coli strain BL21 (DE3); gene DJ1B, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of HIs-tagged enzyme in Escherichia coli strain BL21 (DE3); gene DJ1C, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of HIs-tagged enzyme in Escherichia coli strain BL21 (DE3); gene DJ1E, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of HIs-tagged enzyme in Escherichia coli strain BL21 (DE3); gene DJ1F, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of HIs-tagged enzyme in Escherichia coli strain BL21 (DE3)
gene hchA
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gene PARK7, sequence comparisons
gene ScHSP31 encoding Hsp31 protein, sequence comparisons, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant overexpression of Schizosaccharomyces pombe hsp3101 in Schizosaccharomyces pombe wild-type cells or GLO1 deletion cells; gene ScHSP31, expression in Escherichia coli strain BL21(DE3), detailed phylogenetic analysis of GLOIII, DJ-1 and HSP31 proteins, overview. Overexpression of ScHSP31 can confer methylglyoxal and glyoxal resistance on either wild-type Schizosaccharomyces pombe cells or GLO1-deletion of Schizosaccharomyces pombe
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
glyoxalase III is a stationary-phase enzyme. Its activity reaches a maximum at the entry into the stationary phase and remained high for at least 20 h. Glyoxalase III is regulated by rpoS
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glyoxalase III is not elevated in Escherichia coli cells deleted for glyoxalase I
no induction of glyoxalase III by growth in the presence of methylglyoxal. Paraquat, which can increase the aerobic production of superoxide, suppresses glyoxalase III in JI132
the enzyme is heat-inducible
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C185A
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mutation almost completely abolishes glyoxalase activity
E77A
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mutation almost completely abolishes glyoxalase activity
H184A
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kcat/KM is 3.6fold lower compared to the wild-type value
H186A
-
mutant enzyme shows approximately 17% remaining activity
C111A
site-directed mutagenesis, catalytically inactive mutant
E16A
site-directed mutagenesis, catalytically inactive mutant
H130A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
additional information
Show AA Sequence (288 entries)
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