Information on EC 4.2.1.130 - D-lactate dehydratase

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The expected taxonomic range for this enzyme is: Escherichia coli

EC NUMBER
COMMENTARY
4.2.1.130
-
RECOMMENDED NAME
GeneOntology No.
D-lactate dehydratase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
(R)-lactate = methylglyoxal + H2O
show the reaction diagram
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
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 ACCESSION NO.
COMMENTARY
LITERATURE
glyoxalase III
-
-
glyoxylase III
-
-
-
-
glyoxylase III
-
-
hchA
-
gene name
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Escherichia coli AB1157
-
-
-
Manually annotated by BRENDA team
Escherichia coli MJF274
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
the stationary-phase Escherichia coli cells becomes more susceptible to methylglyoxal when hchA is deleted
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
-
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
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
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
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
-
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 ACCESSION NO.
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
-
Hsp31 efficiently detoxifies exogenously added methylglyoxal
-
-
?
methylglyoxal + H2O
(R)-lactate
show the reaction diagram
Escherichia coli AB1157
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
enhances activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.095
-
Methylglyoxal
-
pH 7.5, temperature not specified in the publication, mutant enzyme H184A
2.61
-
Methylglyoxal
-
pH 7.5, temperature not specified in the publication, wild-type enzyme
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.5
-
Methylglyoxal
-
pH 7.5, temperature not specified in the publication, mutant enzyme H184A
13124
1.8
-
Methylglyoxal
-
pH 7.5, temperature not specified in the publication, wild-type enzyme
13124
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.254
-
-
pH 7.5, temperature not specified in the publication
37
-
-
pH 8.0, 37C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
8
-
no sharp optimum
7.5
-
-
assay at
8
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
82000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 40000, SDS-PAGE
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DTT and either sucrose or mannitol are required for stability
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-15C, 1 mM DTT and 10% sucrose, almost completely stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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 not elevated in Escherichia coli cells deleted for glyoxalase I
-
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 ACCESSION NO.
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
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