Information on EC 1.4.1.8 - valine dehydrogenase (NADP+)

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

EC NUMBER
COMMENTARY hide
1.4.1.8
-
RECOMMENDED NAME
GeneOntology No.
valine dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-valine + H2O + NADP+ = 3-methyl-2-oxobutanoate + NH3 + NADPH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amination
-
-
-
-
Deamination
-
-
-
-
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-valine:NADP+ oxidoreductase (deaminating)
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CAS REGISTRY NUMBER
COMMENTARY hide
37255-39-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC 39118
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strains HK 709, HK706, HK 707, HK 708; VDH activity detectable
-
-
Manually annotated by BRENDA team
strains HK 701, HK 702, HK 703, HK 704, HK 705; VDH activity detectable
-
-
Manually annotated by BRENDA team
Plantago sp.
-
-
-
Manually annotated by BRENDA team
strain 50/137
-
-
Manually annotated by BRENDA team
strain A3
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-2-aminobutyrate + H2O + NAD+
2-oxobutyrate + NADH + NH3
show the reaction diagram
L-alanine + H2O + NAD+
pyruvate + NH3 + NADH
show the reaction diagram
L-cysteine + H2O + NAD+
3-mercapto-2-oxopropanoate + NH3 + NADH
show the reaction diagram
L-isoleucine + H2O + NAD+
3-methyl-2-oxopentanoate + NADH + NH3
show the reaction diagram
L-leucine + H2O + NAD+
2-oxoisocaproate + NADH + NH3
show the reaction diagram
L-methionine + H2O + NAD+
4-(methylthio)-2-oxobutanoic acid + NH3 + NADH
show the reaction diagram
-
VDH2: 2% of the activity with L-valine
-
-
r
L-norleucine + H2O + NAD+
2-oxocaproate + NADH + NH3
show the reaction diagram
L-norvaline + H2O + NAD+
2-oxovalerate + NH3 + NADH
show the reaction diagram
L-phenylalanine + H2O + NAD+
2-oxo-3-phenylpropanoic acid + NH3 + NADH
show the reaction diagram
L-valine + NAD(P)+ + H2O
2-oxoisovalerate + NH3 + NAD(P)H
show the reaction diagram
L-valine + NAD+ + H2O
2-oxoisovalerate + NH3 + NADH
show the reaction diagram
-
-
-
-
?
additional information
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
L-valine + NAD(P)+ + H2O
2-oxoisovalerate + NH3 + NAD(P)H
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,N6-etheno-NAD+
3-acetylpyridine-NAD+
3-pyridinealdehyde-NAD+
Alpha-NAD+
deamino-NAD+
NADP+
NADPH
nicotinamide guanine dinucleotide
additional information
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
2-oxoisovalerate
-
65 mM, 50% loss of specific activity
Ag+
-
0.01 mM in assay buffer
Ca2+
-
1 mM, partial inhibition
diethyldicarbonate
-
20 mM, 60% loss of activity
iodoacetamide
L-valine
Mg2+
-
1 mM, partial inhibition
N-bromosuccinimide
-
0.5 mM, complete inhibition
oxo(phenyl)acetaldehyde
-
5 mM, 60% loss of activity
p-chloromercuribenzoate
p-hydroxymercuribenzoate
pyridoxal 5'-phosphate
-
40% loss of activity, forms Schiff base with a lysine residue
Urea
-
3.5 M, complete and irreversible inactivation
Zn2+
-
1 mM, partial inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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presence of 2-mercaptoethanol or dithiothreitol at 10 mM required for maximal activity
dithiothreitol
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presence of 2-mercaptoethanol or dithiothreitol at 10 mM required for maximal activity
L-valine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.06 - 6.7
2-oxo-3-methylvalerate
0.13 - 1.5
2-oxobutyrate
0.81 - 2.23
2-oxocaproate
0.5 - 1.2
2-oxoisocaproate
0.31 - 50
2-oxoisovalerate
0.48 - 1.94
2-oxovalerate
3.07 - 19.6
L-2-aminobutyrate
28.2 - 333.6
L-alanine
1.18 - 10.8
L-isoleucine
0.67 - 6.3
L-leucine
117.6
L-methionine
-
VDH2
6.68 - 30
L-norleucine
1.33 - 5.7
L-norvaline
0.43 - 714
L-valine
0.029 - 0.43
NAD+
0.022 - 0.093
NADH
0.235
NADP+
-
shoots
0.14
NADPH
-
shoots
15.5 - 153
NH4+
12.3 - 25
pyruvate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
99
2-oxobutyrate
Streptomyces cinnamonensis
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-
107
2-oxocaproate
Streptomyces cinnamonensis
-
-
71.9
2-oxoisocaproate
Streptomyces cinnamonensis
-
-
7.22 - 315
2-oxoisovalerate
164
2-oxovalerate
Streptomyces cinnamonensis
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-
104
L-2-aminobutyrate
Streptomyces cinnamonensis
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-
5.75
L-isoleucine
Streptomyces cinnamonensis
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-
11.9
L-leucine
Streptomyces cinnamonensis
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-
17.9
L-norleucine
Streptomyces cinnamonensis
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-
18
L-norvaline
Streptomyces cinnamonensis
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-
0.000483 - 47.5
L-valine
114
pyruvate
Streptomyces cinnamonensis
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-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
36.8
L-valine
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-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00534
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L-phenylalanine
1.056
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L-cysteine
3.8
-
L-isoleucine
4.2
-
L-leucine
7.02
-
L-isoleucine
7.68
-
L-alanine
19
-
L-2-aminobutyrate
19.56
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oxidative deamination
21.16
-
L-valine
40.92
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reductive amination
52.01
-
2-aminobutyrate
56.33
-
L-valine
64.31
-
oxidative deamination
81.8
-
2-oxoisocaproate
107.6
-
2-oxo-3-methylvalerate
278
-
2-oxobutyrate
302.4
-
2-oxoisovalerate
366.5
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reductive amination
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.5
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reductive amination of 2-oxoisovalerate, 2-oxoisocaproate, 2-oxobutyrate, and DL-2-oxo-3-methylvalerate
8
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reductive amination of 2-oxoisovalerate
8.4 - 8.6
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shoots, reductive amination of valine ketoanalogue
8.5
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reductive amination
8.7 - 8.9
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reductive amination of 2-oxoisovalerate, in presence of 0.3 M Tris-HCl buffer
8.8
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VDH2: reductive amination of 2-oxoisovalerate, 0.3 M Tris-HCl buffer
9.3
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roots, reductive amination of valine ketoanalogue
9.4 - 9.6
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shoots, oxidative deamination of L-valine
10 - 10.5
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oxidative deamination of L-valine, L-norvaline, L-leucine, and L-norleucine
10
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oxidative deamination
10.5 - 10.7
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oxidative deamination of L-valine, in presence of 0.3 M glycine-KCl-KOH buffer
10.7
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oxidative deamination of L-valine, glycine-KCl-KOH buffer
11
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roots, oxidative deamination of L-valine
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5 - 9.5
-
pH 8.5: 40% loss of activity, pH 9.5: 10% loss of activity
10.5
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above rates of reductive amination and oxidative deamination decrease dramatically
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
40
-
enzyme in shoots
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
67000
-
gel filtration
70000
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gel filtration
80000
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VDH2, gel filtration
88000
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gel filtration
116000
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equilibrium ultracentrifugation
118000
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gel filtration
215000
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equilibrium ultracentrifugation
218000
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gel filtration
240000
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VDH2, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 38000, SDS-PAGE
dodecamer
hexamer
tetramer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.3
-
VDH2: stable at pH 6.3 during purification step Reactive-Blue 2 Sepharose chromatography
391703
7
-
below markedly unstable
391694
7.4
-
most stable
391694
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
0.75 M ammonia buffer pH 8.5, half-life: 12 days
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol stabilizes
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.2 M Tris-HCl buffer, pH 7.4, 3 months, no loss of activity
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-20C, 50 mM Tris-HCl buffer, pH 7.4, 5 mM 2-mercaptoethanol, 1 mM EDTA, 30% v/v glycerol, 24 h, 90% loss of activity
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-20C, 70% (NH4)2SO4 suspension in 0.2 M Tris-HCl buffer, 5 months, stable
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-25C, 0.2 M Tris-HCl buffer, pH 7.4, 2 months, no loss of activity
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-70C, 25 mM sodium phosphate buffer pH 7, equal volume of glycerol, 3 months, stable
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-80C, 100 mM Tris-HCl buffer, pH 7.4, 20% v/v glycerol, 2 months, stable
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-80C, 50 mM potassium phosphate pH 7.5, 10% v/v glycerol, 1 mM EDTA, 5 mM 2-mercaptoethanol, 1 month, loss of specific activity
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4C, 0.2 M Tris-HCl buffer, pH 7.4, 24 h, no loss of activity
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4C, 50 mM Tris-HCl buffer, pH 7.4, 5 mM 2-mercaptoethanol, 1 mM EDTA, 10% v/v glycerol, 48 h, stable
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4C, 70% (NH4)2SO4 suspension, 3 weeks, no loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
143fold purification
-
1508fold purification
-
180fold purification
-
21fold purification
-
785fold purification
-
His-bind resin column, 95% purity
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shoots: 21fold purification, roots: 3.9fold purification
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soluble enzyme from roots, 3.9fold purification
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VDH and A124G mutant, His-bind resin column
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VDH2, 346fold purification
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VDH2, 52.6fold purification
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
valDH gene cloned to identify the active site, coding region expressed in Escherichia coli using a pET expression system
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vdh gene
vdh gene encoding VDH2, only one gene responsible for VDH activity, VDH2 is the only active VDH
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A124G
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lower activity toward aliphatic amino acid substrates, e.g. catalytic rate constant for oxidative deamination of L-valine is only 12% of the wild type, but higher activities toward L-phenylalanine, L-tyrosine, and L-methionine
C251S
-
kcat for deamination is 13% of the wild-type value
C30S
-
kcat for deamination is 13% of the wild-type value
K79A
-
lower affinity for L-valine and 2-oxoisovalerate and lower catalytic rate constants compared to the wild type enzyme
K91A
-
lower affinity for L-valine and 2-oxoisovalerate and lower catalytic rate constants compared to the wild type enzyme
V305L
-
kcat for deamination is 0.1% of the wild-type value
additional information
-
active site lysine residue modified with pyridoxal 5-phosphate