Information on EC 1.1.1.45 - L-gulonate 3-dehydrogenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.1.45
-
RECOMMENDED NAME
GeneOntology No.
L-gulonate 3-dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-gulonate + NAD+ = 3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
D-glucuronate degradation I
-
-
Pentose and glucuronate interconversions
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
L-gulonate:NAD+ 3-oxidoreductase
Also oxidizes other L-3-hydroxyacids.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-51-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
sheep
-
-
Manually annotated by BRENDA team
rat
-
-
Manually annotated by BRENDA team
hog
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
L-gulonate 3-dehydrogenase is a bifunctional dimeric protein that functions not only as an NAD+-dependent enzyme in the uronate cycle but also as a taxon-specific lambda-crystallin in rabbit lens
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-keto-L-gulonate + NADH
2,3-diketo-L-gulonate + NAD+
show the reaction diagram
-
-
-
-
ir
? + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
?
acetoacetate + NADH
? + NAD+
show the reaction diagram
-
-
-
-
?
D-galactonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
D-gluconic acid + NAD+
?
show the reaction diagram
D-lyxonic acid + NAD+
?
show the reaction diagram
D-mannonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
D-talonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
D-xylonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
L-3-hydroxybutanoate + NADH
acetoacetate + NAD+
show the reaction diagram
L-3-hydroxybutyrate + ?
?
show the reaction diagram
-
-
-
-
?
L-altronic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
L-erythronic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
L-gulonate + NAD+
3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
L-gulonate + NADP+
3-dehydro-L-gulonate + NADPH + H+
show the reaction diagram
NAD+ is preferred as cofactor
-
-
r
L-gulonic acid + NAD+
3-dehydro-L-gulonic acid + NADH + H+
show the reaction diagram
L-idonic acid + NAD+
?
show the reaction diagram
L-ribonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
L-threonate + ?
?
show the reaction diagram
-
-
-
-
?
L-threonic acid + NAD+
?
show the reaction diagram
-
-
-
-
-
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
L-gulonate + NAD+
3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
P14755
-
-
-
r
L-gulonic acid + NAD+
3-dehydro-L-gulonic acid + NADH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
cofactor binding mode, overview
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no activation or inhibition Zn2+, Cd2+, Cu2+, Fe3+, Mn2+, Ni2+, Co3+, As3+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetoacetate
-
competitive inhibitor
Cibacron blue
-
competitive inhibition with respect to NAD+
malonate
-
reversible inhibition, IC50: 0.34 mM
malonic acid monoethyl ester
-
weak inhibition, IC50: 5.5 mM
NADH
-
competitive inhibitor
p-chloromercuribenzoate
p-Chloromercuriphenylsulfonate
-
0.001 mM, complete inhibition
Phenylmercurinitrate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cysteine
-
in mM-concentration, no activation of the reverse reaction
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.3
2,3-diketo-L-gulonate
-
cofactor NADH, pH 6.3
6.4 - 9.9
acetoacetate
0.45
beta-keto-L-gulonate
-
cofactor NADH, pH 6.3
19
D-gluconate
-
cofactor NAD+, pH 8.5
5.3 - 22.8
L-3-hydroxybutanoate
2.2 - 2.3
L-3-hydroxybutyrate
0.018 - 67
L-gulonate
5.3
L-Idonate
-
cofactor NAD+, pH 8.5
1.2 - 2.9
L-threonate
0.01 - 2.6
NAD+
0.0005 - 0.0006
NADH
0.12 - 0.89
NADP+
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.733
DL-3-hydroxybutanoate
Drosophila melanogaster
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-
7.4
L-gulonate
Drosophila melanogaster
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-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.34
malonate
Oryctolagus cuniculus
-
reversible inhibition, IC50: 0.34 mM
5.5
malonic acid monoethyl ester
Oryctolagus cuniculus
-
weak inhibition, IC50: 5.5 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.011
-
in the supernatant
6.71
-
after purification
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3
-
reduction of 3-dehydro-L-gulonic acid
8.5
-
-
9
-
oxidation of L-gulonic acid
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 8
-
reduction of acetoacetate
6.2 - 10
-
oxidation of L-3-hydroxybutanoate
7 - 10
-
oxidation of 3-hydroxybutanoate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
-
SDS-PAGE
60000 - 70000
63000
-
sucrose density gradient centrifugation
63500
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SDS-PAGE
66000 - 70000
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33000/unit in vitro, 35000/unit in vivo, SDS-PAGE
70000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
X-ray diffraction, natural active protein is presumably a dimer 2 * 36000
additional information
the enzyme possesses two domains, the N-terminal domain with a Rossmann fold and the C-terminal domain with a helical fold. In the N-terminal domain of the NADH-bound structure exist 11 coenzyme-binding residues and two distinct side-chain conformers of coenzyme binding residue Ser124. Subunit dimerization is mediated by numerous intersubunit interactions, including 22 hydrogen bonds and 104 residue pairs of van der Waals interactions, of which those between two cognate C-terminal domains are predominant. Domain and subunit interface structures, structure comparison with the human enzyme, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no modification
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-
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme in both apo form and NADH-bound holo form, X-ray diffraction structure determination and analysis at 1.70-1.85 A resolution
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
in solution instable, by addition of below mM of cyanide several days in the cold, minor effect through EDTA-addition
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
3 M urea results in loss of activity of 83% in 7 min
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18C, 0.05 M K2HPO4, several weeks, little loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate fractionation, Sephadex G-100 column chromatography, Q-Sepharose column chromatography, Blue-Sepharose column chromatography, hydroxylapatite column chromatography
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recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) cells
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) cells
in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D36R/Q41N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E157Q
-
mutation produces no large kinetic alterations
E97Q
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
H145Q
-
inactive enzyme form
N195D
-
inactive enzyme form
N196Q
-
inactive enzyme form
Q41N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S124A
-
mutation decreases the catalytic efficiency 500 fold
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine