Information on EC 1.1.1.B20 - meso-2,3-butandiol dehydrogenase

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

EC NUMBER
COMMENTARY hide
1.1.1.B20
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
meso-2,3-butandiol dehydrogenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2R,3S)-butane-2,3-diol + NAD+ = acetoin + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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SYSTEMATIC NAME
IUBMB Comments
(2R,3S)-butane-2,3-diol:NAD+ oxidoreductase
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
isolated from soil samples
UniProt
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
gene budC
-
-
Manually annotated by BRENDA team
no activity in Escherichia coli
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-
-
Manually annotated by BRENDA team
gene budC
UniProt
Manually annotated by BRENDA team
gene budC
UniProt
Manually annotated by BRENDA team
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AEF50077
GenBank
Manually annotated by BRENDA team
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AEF50077
GenBank
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
-
pathways for the synthesis of 2,3-butanediol in bacteria, overview
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
(2R,3S)-butane-2,3-diol + NAD+
(3R)-acetoin + NADH + H+
show the reaction diagram
(2R,3S)-butane-2,3-diol + NAD+
acetoin + NADH + H+
show the reaction diagram
(2S,3S)-2,3-butanediol + NAD+
(3S)-acetoin + NADH
show the reaction diagram
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(2S,3S)-2,3-butanediol is a poor substrate
-
-
?
(2S,3S)-2,3-butanediol + NAD+
(3S)-acetoin + NADH + H+
show the reaction diagram
(3R)-acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
(3S)-acetoin + NADH + H+
(2S,3S)-2,3-butanediol + NAD+
show the reaction diagram
(R)-acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
(R)-acetoin + NADPH + H+
meso-2,3-butanediol + NADP+
show the reaction diagram
(S)-acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
-
-
-
-
r
1,2-pentanediol + NADH + H+
?
show the reaction diagram
1,2-propandiol + NADH + H+
?
show the reaction diagram
low activity
-
-
?
2 (R,S)-acetoin + 2 NADPH + 2 H+
(2S,3S)-2,3-butanediol + meso-2,3-butanediol + NADP+
show the reaction diagram
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Ara1p is selective toward the acetoin carbonyl group, leading to an S-alcohol
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r
2 acetoin + 2 NADH + 2 H+
(2S,3S)-2,3-butanediol + meso-2,3-butanediol + 2 NAD+
show the reaction diagram
-
racemic mixture of (3S/3R)-acetoin
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-
r
4-methyl-2-pentanone + NADH + H+
4-methyl-2-pentanol + NAD+
show the reaction diagram
acetoin + NADH + H+
(2R,3S)-butane-2,3-diol + NAD+
show the reaction diagram
acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
diacetyl + NADH + H+
(3S)-acetoin + NAD+
show the reaction diagram
-
-
-
?
glycerol + NADH + H+
?
show the reaction diagram
low activity
-
-
?
L-acetoin + NADH
L-2,3-butanediol + NAD+
show the reaction diagram
-
-
-
?
meso-2,3-butanediol + NAD+
(3R)-acetoin + NADH + H+
show the reaction diagram
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-
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r
meso-2,3-butanediol + NAD+
(R)-acetoin + NADH + H+
show the reaction diagram
meso-2,3-butanediol + NAD+
acetoin + NADH
show the reaction diagram
-
-
-
-
?
meso-2,3-butanediol + NAD+
acetoin + NADH + H+
show the reaction diagram
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-
-
-
?
meso-2,3-butanediol + NADP+
(R)-acetoin + NADPH + H+
show the reaction diagram
very low activity
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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
(2R,3S)-butane-2,3-diol + NAD+
(3R)-acetoin + NADH + H+
show the reaction diagram
(2R,3S)-butane-2,3-diol + NAD+
acetoin + NADH + H+
show the reaction diagram
(3R)-acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
(R)-acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
acetoin + NADH + H+
(2R,3S)-butane-2,3-diol + NAD+
show the reaction diagram
acetoin + NADH + H+
meso-2,3-butanediol + NAD+
show the reaction diagram
meso-2,3-butanediol + NAD+
(R)-acetoin + NADH + H+
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
enhances for oxidation and reduction reactions
Mg2+
enhances diacetyl and (3S/3R)-acetoin reduction
Mn2+
enhances diacetyl and (3S/3R)-acetoin reduction
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3S)-2,3-butanediol
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product inhibition
2-mercaptoethanol
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competitive
Fe3+
inhibits diacetyl and (3S/3R)-acetoin reduction
additional information
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no inhibition by NAD+ at 2 mM and 5 mM
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.052 - 31.2
(2S,3S)-2,3-butanediol
0.65 - 0.97
(3R)-acetoin
1.49
(R)-acetoin
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pH 7.0, 30C, recombinant enzyme
6
4-methyl-2-pentanone
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pH 8.0-9.0, 35C, recombinant enzyme
0.23
acetoin
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racemic (R/S)-acetoin, pH 7.0, 30C, recombinant enzyme
3.3
diacetyl
pH 8.0, 40C
2 - 13
meso-2,3-butanediol
0.66
NAD+
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pH 8.0-9.0, 35C, recombinant enzyme
0.05
NADH
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pH 8.0-9.0, 35C, recombinant enzyme
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.02
(2S,3S)-2,3-butanediol
pH 5.0, 40C
19.7
(3R)-acetoin
pH 8.0, 40C
11.5
diacetyl
pH 8.0, 40C
6.2
meso-2,3-butanediol
pH 5.0, 40C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.52
2-mercaptoethanol
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pH 8.0, temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.3
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recombinant Escherichia coli strain BL21(DE3) expressing wild-type BDH, pH 7.0, 30C
1.1
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mutant N146F, substrate (2S,3S)-2,3-butanediol, pH 8.0, 37C
2.17
AEF50077
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh-nox-vgb, pH 8.0, 22C
2.37
purified recombinant enzyme, pH 7.6, 37C, substrates meso-2,3-butanediol and NADP+
2.47
AEF50077
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh-nox, pH 8.0, 22C
3.1
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mutant Q140I/N146F, substrate (2,3S)-2,3-butanediol, pH 8.0, 37C
3.32
AEF50077
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh, pH 8.0, 22C
4.5
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mutant Q140I/N146F, substrate meso-2,3-butanediol, pH 8.0, 37C
5.5
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wild-type, substrate (2S,3S)-2,3-butanediol, pH 8.0, 37C
7.14
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crude recombinant enzyme, pH 7.0, 37C
19.4
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mutant N146F, substrate meso-2,3-butanediol, pH 8.0, 37C
21
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purified reconstituted, recombinant enzyme, reduction of 4-methyl-2-pentanone, pH 8.0, 35C
66
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purified reconstituted, recombinant enzyme, oxidation of meso-2,3-butanediol, pH 8.0, 35C
119.55
purified recombinant enzyme, pH 7.6, 37C, substrates meso-2,3-butanediol and NAD+
177
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wild-type, substrate meso-2,3-butanediol, pH 8.0, 37C
218
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purified reconstituted, recombinant enzyme, reduction of acetoin, pH 8.0, 35C
461.67
purified recombinant enzyme, pH 7.6, 37C, substrates acetoin and NADH
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
maximal activity of (3S/3R)-acetoin reduction
10.5
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2,3-butanediol oxidation assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
AEF50077
in vitro assay at room temperature
30 - 37
AEF50077
in vivo assay at
40
optimum for oxidation and reduction reactions
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in complex with substrate meso-2,3-butanediol and inhibitor 2-mercaptoethanol, to 1.7 A resolution. The overall strucuture is similar to that of other short chain dehydrogenase/reductase enzymes, the NAD+ binding site, and the positions of catalytic residues Ser139, Tyr152, and Lys156 are also conserved. 2-Mercaptoethanol forms hydrogens bonds with residues Gln 140 and Gly 183 close to the active site, which are important but not sufficient for distiguishing stereoisomerism of a chiral substrate
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
lyophilised recombinant ADH-9 from Escherichia coli BL21(DE3) cells by immobilization onto Amberlite FPA54 to 0.01 U per mg carrier
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ADH-9 gene is identified from a biodiversity library, recombinant expression in Escherichia coli strain BL21/(DE3)
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expression in Escherichia coli
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gene bdh, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, functional recombinant expression in Escherichia coli
gene bdh1, phylogenetic analysis, expression in Escherichia coi strain YYC202(DE3) ldhA-/- ilvC? expressing ilvBN from Escherichia coli and aldB from Lactobacillus lactis, encoding acetolactate synthase and acetolactate decarboxylase activities, respectively, disruption of the lactate biosynthesis pathway in the strain increases pyruvate precursor availability to this strain, increased availability of NADH for acetoin reduction to meso-2,3-butanediol is the most important consequence of ldhA deletion
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gene budC , recombinant expression in Escherichia coli strain BL21(DE3)
gene budC, expression in Escherichia coli strain BL21(DE3) pLys
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gene budC, phylogenetic tree, functional recombinant expression in Bacillus subtilis strain 168
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gene budC, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
gene budC, sequence comparisons, expression in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha
gene butACg, subcloning in Escherichia coli strain DH5alpha, recombinant expression in Escherichia coli strain BL21(DE3), coexpression with the genes encoding ALS and ALDC of Lactobacillus lactis strain MG1363
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gene mbdh, functional coexpression of Serratia sp. meso-2,3-butanediol dehydrogenase, Lactobacillus brevis NADH oxidase and Vitreoscilla sp. hemoglobin in Escherichia coli strain BL21(DE3)/pET-mbdh-nox-vgb, construction of plasmids pET-mbdh, pET-mbdh-nox, and pET-mbdh-nox-vgb, method optimization
AEF50077
phylogenetic analysis, enzyme expression in Escherichia coli strains MG1655(DE3) and YYC202(DE3)
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D194G
site-directed mutagenesis, the mutant binds the substrate but is catalytically almost inactive. The mutant is inactive with (2S,3S)-butanediol, meso-butanediol and (2R,3R)-butanediol. D194G enzyme mutant shows a similar secondary structure compared to Enterobacter aerogenes BDH. While the mutant is highly susceptible to protease digestion compared to the wild-type enzyme. Homology modeling of the mutant enzyme, with meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae, PDB ID 1GEG, as a template, reveals that Gly194 seems to lose the hydrogen bond interactions with the surrounding residues (Gly206, Gly207 and Thr209), resulting in a putative conformational changes of mutant D194G which might be responsible for the loss of activity
moe
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construction and engineering of Corynebacterium glutamicum strain DELTAaceEDELTApqoDELTAldhA(pEKEx2-als,aldB,butACg). Chromosomal inactivation of the putative BDH gene butACg (cg2958) in strain DELTAaceEDELTApqoDELTAldhA. BDH activity is nearly abolished upon inactivation of butACg indicating that Corynebacterium glutamicum expresses a single BDH under the experimental conditions examined. BDH activity increases 3fold in strain DELTAaceEDELTApqoDELTAldhA(pEKEx2-als,aldB,butACg) compared to the respective control. The inactivation of butACg gene decreases the BDH activity 75fold for the DELTAaceEDELTApqoDELTAldhADELTAbutACg(pEKEx2) strain compared to strain DELTAaceEDELTApqoDELTAldhA(pEKEx2). The major form of 2,3-butanediol is meso-2,3-butandediol, and the ratio meso-2,3-BD/optically active 2,3-BD is 95:5, the main side products are glycerol, ethanol, and acetoin
N146F
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11% of wild-type specific activity
Q140I
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trace activity below 0.1 U/mg
Q140I/N146F
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poor activity
Q140I/N146F/W190H
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trace activity below 0.1 U/mg with substrate meso-butanediol, 2.9 U/mg with substrate (2S,3S)-2,3-butanediol
Q140I
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mutation mimicking the corresponding residue in (S,S)-butanediol dehydrogenase. No activity with substrates meso-butanediol or (S,S)-butanediol
Q140I/N146F
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mutation mimicking the corresponding residues in (S,S)-butanediol dehydrogenase. Poor activity with substrates meso-butanediol or (S,S)-butanediol
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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discovery of the (S)-selective alcohol dehydrogenase enables a novel production process of (R)-acetoin from meso-2,3-butanediol