Information on EC 4.2.1.130 - D-lactate dehydratase

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

EC NUMBER
COMMENTARY
4.2.1.130
-
RECOMMENDED NAME
GeneOntology No.
D-lactate dehydratase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(R)-lactate = methylglyoxal + H2O
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
methylglyoxal degradation
-
-
methylglyoxal degradation II
-
-
Microbial metabolism in diverse environments
-
-
Pyruvate metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
(R)-lactate hydro-lyase
The enzyme converts methylglyoxal to D-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).
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glutathione-independent glyoxalase III
Q04432
-
glutathione-independent glyoxalase III
Q10356
-
glyoxalase III
-
-
glyoxalase III
Q04432
-
glyoxalase III
Q10356
-
glyoxylase III
-
-
-
-
glyoxylase III
-
-
GSH-independent glyoxalase III
Q04432
-
GSH-independent glyoxalase III
Q10356
-
hchA
-
gene name
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Escherichia coli AB1157
-
-
-
Manually annotated by BRENDA team
Escherichia coli MJF274
-
-
-
Manually annotated by BRENDA team
gene ScHSP31
UniProt
Manually annotated by BRENDA team
genes hsp3101 and hsp3102
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
detailed phylogenetic analysis of GLOIII, DJ-1 and HSP31 proteins in fungi, overview
evolution
Q04432
detailed phylogenetic analysis of GLOIII, DJ-1 and HSP31 proteins in fungi, overview
physiological function
-
glutathione-dependent glyoxalase pathway, i.e. glyoxalase I/II, is the most important route for the in vivo detoxification of methylglyoxal. Glyoxalase III may play a critical role in conditions with limiting carbon source. The enzyme may play an important role in protecting stationary-phase cells against carbonyl toxicity
physiological function
-
in enteric bacteria methylglyoxal is detoxified by the glutathione-dependent glyoxalase I/II system, by glyoxalase III, and by aldehyde reductase and alcohol dehydrogenase. Glyoxalase III might be important for survival of non-growing Escherichia coli cultures
physiological function
-
the defensive glyoxalase III is inactivated by the oxidative stress imposed by the lack of superoxide dismutase, thereby exacerbating the deleterious effect of sugar oxidation
physiological function
-
glyoxalase III is responsible for the detoxification of reactive carbonyl species, e.g. methylglyoxal and glyoxal, via the GSH-independent pathway, overview
physiological function
Q04432
glyoxalase III is responsible for the detoxification of reactive carbonyl species, e.g. methylglyoxal and glyoxal, via the GSH-independent pathway, overview
physiological function
Escherichia coli AB1157
-
the defensive glyoxalase III is inactivated by the oxidative stress imposed by the lack of superoxide dismutase, thereby exacerbating the deleterious effect of sugar oxidation
-
malfunction
-
the stationary-phase Escherichia coli cells becomes more susceptible to methylglyoxal when hchA is deleted
additional information
-
two Schizosaccharomyces pombe Hsp31 proteins (Hsp3101 and Hsp3102) and one Saccharomyces cerevisiae Hsp31 protein (ScHsp31) display significantly higher in vitro GLO3 activity than Schizosaccharomyces pombe DJ-1 (SpDJ-1)
additional information
Q04432
two Schizosaccharomyces pombe Hsp31 proteins (Hsp3101 and Hsp3102) and one Saccharomyces cerevisiae Hsp31 protein (ScHsp31) display significantly higher in vitro GLO3 activity than Schizosaccharomyces pombe DJ-1 (SpDJ-1)
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
-
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Q10356
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Q04432
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
-
Hsp31 efficiently detoxifies exogenously added methylglyoxal
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
-
glyoxalase III does not catalyse the reverse reaction. Glyoxalase III cannot catalyse either the formation or the breakdown of S-D-lactoylglutathione
-
-
ir
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Escherichia coli AB1157
-
-
-
-
?
phenylglyoxal + H2O
?
show the reaction diagram
-
the rate with phenylglyoxal is about 15% of that with methylglyoxal
-
-
ir
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
-
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Q10356
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Q04432
-
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
-
Hsp31 efficiently detoxifies exogenously added methylglyoxal
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Escherichia coli AB1157
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe2+
-
enhances activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5,5'-dithiobis-(2-nitrobenzoate)
-
0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
Borate
-
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%
N-ethylmaleimide
-
0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
p-hydroxymercuribenzoate
-
0.4 mM, 15 min, complete inactivation. 5 mM DTT restores activity almost completely
superoxide
-
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+
-
maximal inhibition above 0.3 mM, 10% inhibition at 0.025 mM, more than 50% inhibition at 0.1 mM
hydrogen peroxide
-
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
additional information
-
S-methylglutathione and S-octylglutathione, even at concentrations of 20 mM and 0.25 mM respectively, have no inhibitory effect on Escherichia coli glyoxalase III
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.19
methylglyoxal
-
pH 7.5, temperature not specified in the publication, mutant enzyme H184A
1.43
methylglyoxal
-
pH 7.5, temperature not specified in the publication, wild-type enzyme
1.5
methylglyoxal
Q04432
recombinant ScHsp31, pH and temperature not specified in the publication
10.8
methylglyoxal
Q10356
recombinant wild-type SpDJ-1, pH and temperature not specified in the publication
31.1
methylglyoxal
Q10356
recombinant Hsp3101, pH and temperature not specified in the publication
37.3
methylglyoxal
Q10356
recombinant mutant SpDJ-1 H130A, pH and temperature not specified in the publication
58
methylglyoxal
Q10356
recombinant Hsp3102, pH and temperature not specified in the publication
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.095
methylglyoxal
-
pH 7.5, temperature not specified in the publication, mutant enzyme H184A
0.518
methylglyoxal
Q10356
recombinant Hsp3101, pH and temperature not specified in the publication
0.967
methylglyoxal
Q10356
recombinant Hsp3102, pH and temperature not specified in the publication
1.09
methylglyoxal
Q10356
recombinant mutant SpDJ-1 H130A, pH and temperature not specified in the publication
1.25
methylglyoxal
Q04432
recombinant ScHsp31, pH and temperature not specified in the publication
1.43
methylglyoxal
Q10356
recombinant wild-type SpDJ-1, pH and temperature not specified in the publication
2.61
methylglyoxal
-
pH 7.5, temperature not specified in the publication, wild-type enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.03
methylglyoxal
Q10356
recombinant mutant SpDJ-1 H130A, pH and temperature not specified in the publication
322
0.132
methylglyoxal
Q10356
recombinant wild-type SpDJ-1, pH and temperature not specified in the publication
322
0.35
methylglyoxal
Q10356
recombinant Hsp3102, pH and temperature not specified in the publication
322
0.383
methylglyoxal
Q10356
recombinant Hsp3101, pH and temperature not specified in the publication
322
0.5
methylglyoxal
-
pH 7.5, temperature not specified in the publication, mutant enzyme H184A
322
0.85
methylglyoxal
Q04432
recombinant ScHsp31, pH and temperature not specified in the publication
322
1.8
methylglyoxal
-
pH 7.5, temperature not specified in the publication, wild-type enzyme
322
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.254
-
pH 7.5, temperature not specified in the publication
37
-
pH 8.0, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 8
-
no sharp optimum
7.5
-
assay at
8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 10
-
pH 5.0: sharp decrease in reaction rate below, pH 10.0: activity 20-25% lower than the rate at physiological pH
5 - 9
-
pH 5.0: about 45% of maximal activity, pH 9.0: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25 - 50
-
25C: about 55% of maximal activity, 50C: about 75% of maximal activity
PDB
SCOP
CATH
ORGANISM
Candida albicans (strain SC5314 / ATCC MYA-2876)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
82000
-
gel filtration
714075
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
2 * 40000, SDS-PAGE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
DTT and either sucrose or mannitol are required for stability
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, 1 mM DTT and 10% sucrose, almost completely stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
Q04432
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, 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
Q10356
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
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
-
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
-
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
Escherichia coli AB1157
-
-
glyoxalase III is not elevated in Escherichia coli cells deleted for glyoxalase I
Escherichia coli MJF274
-
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C185A
-
mutation almost completely abolishes glyoxalase activity
E77A
-
mutation almost completely abolishes glyoxalase activity
H184A
-
kcat/KM is 3.6fold lower compared to the wild-type value
C111A
Q10356
site-directed mutagenesis, catalytically inactive mutant
E16A
Q10356
site-directed mutagenesis, catalytically inactive mutant
H186A
-
mutant enzyme shows approximately 17% remaining activity
additional information
-
mutagenesis studies based on evaluation of conserved catalytic residues reveals that the Cys185 and Glu77 are essential for catalysis, whereas His186 is less crucial for enzymatic function, although it participates in the catalytic process
H130A
Q10356
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
additional information
-
overexpression of Saccharomyces cerevisiae gene ScHSP31 can confer methylglyoxal and glyoxal resistance on either wild-type Schizosaccharomyces pombe cells or GLO1-deletion of Schizosaccharomyces pombe