Information on EC 1.1.99.6 - D-lactate dehydrogenase (acceptor)

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

EC NUMBER
COMMENTARY hide
1.1.99.6
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RECOMMENDED NAME
GeneOntology No.
D-lactate dehydrogenase (acceptor)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-lactate + acceptor = pyruvate + reduced acceptor
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
alanine metabolism
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SYSTEMATIC NAME
IUBMB Comments
(R)-lactate:acceptor 2-oxidoreductase
The zinc flavoprotein (FAD) from the archaeon Archaeoglobus fulgidus cannot utilize NAD+, cytochrome c, methylene blue or dimethylnaphthoquinone as acceptors. In vitro it is active with artificial electron acceptors such as 2,6-dichlorophenolindophenol, but the physiological acceptor is not yet known.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-83-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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mutations in the D2HGDH gene, encoding D-2-hydroxyglutarate dehydrogenase, cause D-2-hydroxyglutaric aciduria
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-2-hydroxybutyrate + oxidized 2,6-dichlorophenolindophenol
2-oxobutyrate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
(R)-lactate + acceptor
pyruvate + reduced acceptor
show the reaction diagram
(R)-lactate + ferricyanide
pyruvate + ferrocyanide
show the reaction diagram
(R)-lactate + oxidized 2,6-dichlorophenolindophenol
pyruvate + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium
show the reaction diagram
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-
-
-
?
(R)-lactate + oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
pyruvate + reduced 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
show the reaction diagram
2(R)-hydroxybutanoate + acceptor
2-oxobutanoate + reduced acceptor
show the reaction diagram
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-
-
?
2(R)-hydroxyheptanoate + acceptor
2-oxoheptanoate + reduced acceptor
show the reaction diagram
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-
-
?
2(R)-hydroxyhexanoate + acceptor
2-oxohexanoate + reduced acceptor
show the reaction diagram
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-
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?
2(R)-hydroxyoctanoate + acceptor
2-oxooctanoate + reduced acceptor
show the reaction diagram
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-
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?
2(R)-hydroxypentanoate + acceptor
2-oxopentanoate + reduced acceptor
show the reaction diagram
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?
D-gluconate + acceptor
2-oxo-gluconate + reduced acceptor
show the reaction diagram
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r
D-glucose 6-phosphate + acceptor
?
show the reaction diagram
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r
D-glycerate + acceptor
hydroxypyruvate + reduced acceptor
show the reaction diagram
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-
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?
D-malate + acceptor
oxaloacetate + reduced acceptor
show the reaction diagram
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-
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?
D-tartrate + acceptor
2-oxo-3-hydroxybutanoate + reduced acceptor
show the reaction diagram
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poor substrate
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?
meso-tartrate + acceptor
2-oxo-3-hydroxybutanoate + reduced acceptor
show the reaction diagram
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good substrate
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?
N-acetylneuraminate + acceptor
?
show the reaction diagram
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r
phenylpyruvate + reduced acceptor
phenyl-(R)-lactate + acceptor
show the reaction diagram
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r
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
(R)-lactate + acceptor
pyruvate + reduced acceptor
show the reaction diagram
additional information
?
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O29853
the enzyme is expressed constitutively
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,6-dichlorophenolindophenol
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anthraquinone-2,6-disulfonate
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artificial electron acceptor
cytochrome c
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possible physiological acceptor
molybdopterin mononucleotide
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phenazine methosulfate
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ubiquinone
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possible physiological acceptor
additional information
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molybenum-containing Fe-S-enzyme without an additional prosthetic group such as flavin or cytochrome c, enzyme contains one molybdenum, four iron and four sulfur atoms per subunit
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCN
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raises dehydrogenase activity of freshly prepared kidney extracts
Molybdenum
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molybdenum containing enzyme
Zn2+
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the enzyme contains 1 mol of Zn2+ per mol of protein
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
4-chloromercuribenzoate
1 mM, 62% loss of activity
CoCl2
1 mM, 78% loss of activity
CuCl2
1 mM, complete loss of activity
HgCl2
1 mM, 93% loss of activity
L-lactate
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weak
MnCl2
1 mM, complete loss of activity
oxalate
oxaloacetate
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competitive to acceptor
p-chloromercuribenzoate
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strong
pyruvate
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competitive inhibitor to acceptor
Tris
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slight inhibition above 30 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate
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stimulates
ferricyanide
methylene blue
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n-dodecyl beta-D-maltoside
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stimulates
phenazine methosulfate
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stimulates
Triton X-100
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stimulates
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.091 - 0.156
(R)-lactate
2 - 3.4
D-lactate
1
D-malate
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0.247
oxidized 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
pH 8.5, 60°C, cosubstrate: (R)-lactate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
270
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80000 Da form of the enzyme
1800
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600000 Da form of the enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
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and below for malate as substrate
8
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in a 25 mM Tris buffer at 60°C
8.6
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D-lactate as substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
pH 6.5: about 50% of maximal activity, pH 9.0: about 50% of maximal activity
7 - 9.5
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pH 7.0: about 30% of maximal activit, pH 9.5: about 55% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 95
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25°C: 8% of maximal activity, 95°C: about 75% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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more than 85% of the enzyme is associated with the membrane, integral membrane protein, a significant portion including part of the FAD-binding pocket, is outside the membrane facing the S-layer. NADH oxidase (NoxA2) co-localized to the same sites in the membrane (NoxA2) may protect proteins involved in electron transfer by reducing O2 to H2O2 or H2O
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48000
2 * 48000, SDS-PAGE
49715
2 * 49715, calculated from sequence
80000
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x * 80000, probably octamer, SDS-PAGE
93000
non-denaturing PAGE
102000
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gel filtration
180000 - 600000
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gel filtration, native PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
oligomer
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x * 80000, probably octamer, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour-diffusion method with polyethylene glycol 8000 as the precipitant. The crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 63.4, b = 119.4, c = 70.2 A, beta = 112.0°, and diffract to 2.0 A resolution
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 10
50°C, 30 min, enzyme retains full activity
693285
8
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in 25 mM Tris buffer at 4°C stable for weeks
287840
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
10 min, enzyme retains full activity
83
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half-life: 105 min
85
10 min, more than 60% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
oxalate stabilizes
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OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
oxygen-sensitive enzyme, has to be handled in the presence of detergent and absence of oxygen, if stirred under air at room temperatures 50% activity are lost within 30 min, the residual activity has a half-life of 10 h
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287841
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0°C, as precipitate in ammonium sulfate stable for months
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4°C, in 25 mM Tris buffer, pH 8.0, stable for weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-cellulose, hydroxyapatite, gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression as part of a fusion protein with maltose-binding protein in Escherichia coli
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expression in Escherichia coli
expression in Escherichia coli, gene is localized on chromosome 9q between marker WI-3028 and WI-93330
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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production of pyruvate in an enzyme-membrane reactor