Information on EC 1.1.1.304 - diacetyl reductase [(S)-acetoin forming]

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

EC NUMBER
COMMENTARY hide
1.1.1.304
-
RECOMMENDED NAME
GeneOntology No.
diacetyl reductase [(S)-acetoin forming]
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(S)-acetoin + NAD+ = diacetyl + NADH + H+
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Butanoate metabolism
-
-
pyruvate fermentation to (S)-acetoin
-
-
acetoin degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
(S)-acetoin:NAD+ oxidoreductase
The reaction is catalysed in the reverse direction. This activity is usually associated with butanediol dehydrogenase activity (EC 1.1.1.4 or EC 1.1.1.76). While the butanediol dehydrogenase activity is reversible, diacetyl reductase activity is irreversible. This enzyme has been reported in the bacteria Geobacillus stearothermophilus, Enterobacter aerogenes and Klebsiella pneumoniae [1-3]. Different from EC 1.1.1.303, diacetyl reductase [(R)-acetoin forming].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
bifunctional diacetyl reductase and (R)-2,3-butanediol dehydrogenase
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
gene dar
KC505218
GenBank
Manually annotated by BRENDA team
gene dar
KC505218
GenBank
Manually annotated by BRENDA team
gene adr
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-pentanedione + beta-NADH + H+
L-3-hydroxy-2-pentanone + beta-NAD+
show the reaction diagram
-
-
-
-
ir
2,3-pentanedione + NADH + H+
3-hydroxy-2-pentanone + NAD+
show the reaction diagram
diacetyl + beta-NADH + H+
(S)-acetoin + beta-NAD+
show the reaction diagram
-
86.9% of the activity with pentane-2,3-dione
-
-
ir
diacetyl + NADH + H+
(S)-acetoin + NAD+
show the reaction diagram
diacetyl + NADPH + H+
(S)-acetoin + NADP+
show the reaction diagram
ethyl pyruvate + beta-NADH + H+
? + beta-NAD+
show the reaction diagram
-
38.4% of the activity with pentane-2,3-dione
-
-
ir
ethyl pyruvate + NADH + H+
? + NAD+
show the reaction diagram
-
57.7% of the activity with diacetyl
-
-
?
methyl glyoxal + NADH + H+
? + NAD+
show the reaction diagram
-
11% of the activity with diacetyl
-
-
?
methyl pyruvate + beta-NADH + H+
? + beta-NAD+
show the reaction diagram
-
22.8% of the activity with pentane-2,3-dione
-
-
ir
methyl pyruvate + NADH + H+
? + NAD+
show the reaction diagram
-
49% of the activity with diacetyl
-
-
?
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
diacetyl + NADH + H+
(S)-acetoin + NAD+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
KC505218
dependent on
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
activates by 201.6 to 265.6% at 2 mM
Mn2+
-
activates by 201.6 to 265.6% at 2 mM
Na+
-
activates by 201.6 to 265.6% at 2 mM
additional information
-
addition of EDTA or the cations at 1 mM, such as Na+, K+, Mn2+, Mg2+, and Ca2+, have no significant effect on the activity of ReADR
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-oxoglutarate
-
noncompetitive
acetoin
-
noncompetitive, product inhibition
acetone
-
competitive for diacetyl, uncompetitive for NADH
diacetyl
-
substrate inhibition at concentrations above 80-90 mM
ethyl pyruvate
-
substrate inhibition at concentrations above 80-90 mM
Fe2+
-
inhibits 91.6% at 2 mM
Hexane-2,5-dione
-
noncompetitive
Methyl pyruvate
-
substrate inhibition at concentrations above 80-90 mM
NAD+
-
competitive, product inhibition
Pentane-3-one
-
competitive for diacetyl, uncompetitive for NADH
Zn2+
-
inhibits 94.6% at 2 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DMSO
-
DMSO at a final concentration of 30% v/v added into the assay mixture, increases the activity up to 120% of the control enzyme activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6
2,3-Pentanedione
-
25°C, pH 6.0
1.6 - 19
diacetyl
20 - 24
ethyl pyruvate
75
methyl glyoxal
-
pH 7.0, 25°C
16 - 18
Methyl pyruvate
0.005 - 0.135
NADH
6
pentane-2,3-dione
-
pH 7.0, 25°C
additional information
additional information
-
Michaelis-Menten-type kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
163
diacetyl
Mycobacterium sp.
W8VSK8
pH 7.0, 30°C
110
NADH
Mycobacterium sp.
W8VSK8
pH 7.0, 30°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
36.4
diacetyl
Mycobacterium sp.
W8VSK8
pH 7.0, 30°C
746
8.519 - 210
NADH
8
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
300
diacetyl
-
25°C, pH 6.0
150
ethyl pyruvate
-
25°C, pH 6.0
150
Methyl pyruvate
-
25°C, pH 6.0
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
71.4
-
pH 7.5, 25°C
72.6
KC505218
purified recombinant enzyme, NADPH, pH 6.0, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
diacetyl reduction
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
-
5 min, 60% loss of activity
5
-
5 min, 30% loss of activity
5.1
-
5 min, 20% loss of activity
5.4 - 7.6
-
stable within
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8
isoelectric focusing
5.9 - 7.2
isoelectric focusing
6.8
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
-
2 * 26000, SDS-PAGE
26591
4 * 26591, calculated
26864
-
2 * 26864, sequence calculation
28000
-
2 * 28000, SDS-PAGE
28500
KC505218
4 * 28500, recombinant enzyme, SDS-PAGE
49000
-
gel filtration
61000
-
gel filtration
68000
-
gel filtration
96000
gel filtration
118000
KC505218
recombinant enzyme, gel filtration
150000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
-
1 * 68000, SDS-PAGE
tetramer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8
-
704903
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable to dilution, kept diluted at 0°C for ca. 60 min it will lose 62% of activity. This inactivation is almost completely reversed by the addition of NAD+
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
storage at 0°C in the presence of 20% glycerol, 0.1 mM EDTA, 5 mM 2-mercaptoethanol and 0.6 mM NAD+ in TEA buffer, pH 7.5, half-life of one month
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme by adsorption on diethylaminoethyl–Sepharose and hydrophobic interaction chromatography
-
recombinant enzyme from Escherichia coli strain Rosetta (DE3)
KC505218
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene adr, DNA and amino acid sequence determination and analysis, sequence comparison, expression of His-tagged enzyme in Escherichia coli
-
gene dar, fucntional expression in Escherichia coli strain Rosetta (DE3), resulting in production of S-acetoin with higher than 99.9% optical purity from diacetyl
KC505218
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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