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Information on EC 1.1.1.169 - 2-dehydropantoate 2-reductase and Organism(s) Escherichia coli and UniProt Accession P0A9J4

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Escherichia coli
UNIPROT: P0A9J4 not found.
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
Synonyms
ketopantoate reductase, 2-dehydropantoate 2-reductase, tk-kpr, kpa reductase, ketopantoic acid reductase, conjugated polyketone reductase, tk1968, 2-ketopantoate reductase, tk1968 protein, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ketopantoate reductase
-
2-ketopantoate reductase
-
-
-
-
2-ketopantoic acid reductase
-
-
-
-
2-oxopantoate reductase
-
-
-
-
ketopantoate reductase
ketopantoic acid reductase
-
-
-
-
KPA reductase
-
-
-
-
additional information
KPR belongs to the 6-phosphogluconate dehydrogenase superfamily in the SCOP database
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-pantoate + NADP+ = 2-dehydropantoate + NADPH + H+
show the reaction diagram
molecular catalytic mechanism, substrate and cofactor binding, Asn98, Glu256, and Lys176 are essential, overview
(R)-pantoate + NADP+ = 2-dehydropantoate + NADPH + H+
show the reaction diagram
protein-ligand interactions, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
(R)-pantoate:NADP+ 2-oxidoreductase
-
CAS REGISTRY NUMBER
COMMENTARY hide
37211-74-8
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-dehydropantoate + NADPH
(R)-pantoate + NADP+
show the reaction diagram
ketopantoate + NADPH
pantoate + NADP+
show the reaction diagram
2-dehydropantoate + NADPH + H+
(R)-pantoate + NADP+
show the reaction diagram
-
-
-
-
?
2-oxoisovalerate + NADPH
2-hydroxyvalerate + NADP+
show the reaction diagram
-
low activity
-
-
r
2-oxopantoate + 3'-NADPH
(R)-pantoate + 3'-NADP+
show the reaction diagram
-
-
-
-
r
2-oxopantoate + alpha-NADPH
(R)-pantoate + alpha-NADP+
show the reaction diagram
-
-
-
-
r
2-oxopantoate + beta-NADPH
(R)-pantoate + beta-NADP+
show the reaction diagram
-
highly specific for
-
-
r
2-oxopantoate + NADPH
(R)-pantoate + NADP+
show the reaction diagram
-
part of the pantothenate biosynthesis
-
-
?
2-oxopantoate + thio-NADPH
(R)-pantoate + thio-NADP+
show the reaction diagram
-
-
-
-
r
3-methyl-2-oxo-n-valerate + NADPH
2-hydroxy-3-methyl-n-valerate + NADP+
show the reaction diagram
-
low activity
-
-
r
alpha-ketopantoate + NADPH
D-pantoate + NADP+
show the reaction diagram
ketopantoate + ?
pantothenate + ?
show the reaction diagram
-
-
-
?
ketopantoate + NADPH
pantoate + NADP+
show the reaction diagram
ketopantoic acid + NADPH
pantoic acid + NADP+
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-dehydropantoate + NADPH
(R)-pantoate + NADP+
show the reaction diagram
ketopantoate + NADPH
pantoate + NADP+
show the reaction diagram
2-dehydropantoate + NADPH + H+
(R)-pantoate + NADP+
show the reaction diagram
-
-
-
-
?
2-oxopantoate + NADPH
(R)-pantoate + NADP+
show the reaction diagram
-
part of the pantothenate biosynthesis
-
-
?
alpha-ketopantoate + NADPH
D-pantoate + NADP+
show the reaction diagram
-
pantothenate/coenzyme A biosynthetic pathway
-
r
ketopantoate + NADPH
pantoate + NADP+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
3'-NADPH
-
-
alpha-NADPH
-
-
beta-NADPH
-
preference for beta-NADPH as cofactor
NADP+
NADPH
thio-NADPH
-
-
additional information
-
NAD+/NADH are poor cofactors
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2'-phospho-ADP-ribose
-
competitive
2'-phospho-AMP
-
competitive
ADP
-
competitive
AMP
-
competitive
ATP
-
noncompetitive
NADP+
-
competitive versus NADPH, noncompetitive versus ketopantoate
Pantoate
-
noncompetitive versus ketopantoate and NADPH
phosphate
-
competitive
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bromoethylamine
-
activation of enzyme from K176C mutant
ethylamine
-
activation of enzyme from K176A mutant
formate
-
E256A mutant activity greatly increased, twice as high at pH 7.2 as at pH 5.9
methylamine
-
activation of enzyme from K176A mutant
Propylamine
-
activation of enzyme from K176A mutant
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03
2-dehydropantoate
pH 7.5, 27°C, wild-type enzyme
0.06 - 0.12
ketopantoate
0.004 - 0.02
NADPH
8.7
2-oxoisovalerate
-
pH 7.5, 25°C, recombinant enzyme
0.12
2-oxopantoate
-
pH 7.5, 25°C, recombinant enzyme
0.075
3'-NADPH
-
pH 7.5, 25°C, recombinant enzyme
3.8
3-methyl-2-oxo-n-valerate
-
pH 7.5, 25°C, recombinant enzyme
0.036
alpha-NADPH
-
pH 7.5, 25°C, recombinant enzyme
0.004
beta-NADPH
-
pH 7.5, 25°C, recombinant enzyme
0.038 - 40
ketopantoate
0.007
NADP+
-
+/-0.002
0.002 - 0.016
NADPH
0.26
Pantoate
-
+/-0.04
0.018
thio-NADPH
-
pH 7.5, 25°C, recombinant enzyme
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
25
2-dehydropantoate
pH 7.5, 27°C, wild-type enzyme
8 - 40
ketopantoate
25
NADPH
pH 7.5, 27°C, wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2
2'-phospho-ADP-ribose
-
pH 7.6
0.24
2'-phospho-AMP
-
pH 7.6
1.05
ADP
-
pH 7.6
6.3
AMP
-
pH 7.6
0.61
ATP
-
pH 7.6
27
phosphate
-
pH 7.6
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
7.6
-
assay at
additional information
-
activity decreases with pH increase, reduction of ketopantoic acid
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
pH-dependence of kinetics with different substrates, modeling, overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33000
SDS-PAGE
34000
33000
-
SDS-PAGE, gel filtration
33800
-
calculated from 912 bp coding region
33866
-
1 * 33000 gel filtration, 1 * 33866 elelectrospray mass spectrometry analysis, 1 * 33870 amino acid sequence
33870
-
1 * 33000 gel filtration, 1 * 33866 elelectrospray mass spectrometry analysis, 1 * 33870 amino acid sequence
34000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
SeMet substitution
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
prismic crystals, hanging drop vapor-diffusion technique
purified enzyme with bound NADP+, hanging drop vapour diffusion method, 10-15 mg/ml protein at 4°C is mixed with ketopantoate and NADP+ in a ratio of 5:1 and 2:1, respectively, in 0.1 M sodium acetate, pH 4.0-5.0, with 10%2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.1 A resolution, ternary complex modelling
purified recombinant His6-tagged enzyme in complex with 2'-monophosphoadenosine 5'-diphosphoribose, 4°C, 10-15 mg/ml protein with NADPH and pantoate at a final ligand:protein ratio of 2:1 and 5:1, respectively, mixing with 10% 2-methyl-2,4-pentanediol buffered with 0.1 M sodium acetate pH 4.0-5.0, X-ray diffraction structure determination and analysis at 1.95-2.0 A resolution
purified recombinant His6-tagged enzyme in complex with NADP+ and pantoate, hanging drop vapor-diffusion technique, 20°C, 15-30 mg/ml protein with 2 mM NADP+ and 10 mM pantoate, 0.002 ml of protein solution is mixed with an equal volume of well solution containing 35% v/v dioxane, cryoprotection with 20% v/v 2-methyl-2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.3 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D248A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
E210A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
E256A
K176A
R31A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
S244A
D248A
-
site-directed mutagenesis, wild-type activity
E210A
-
site-directed mutagenesis, wild-type activity
E240A
-
site-directed mutagenesis, wild-type activity
E256A
-
site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme
E256D
-
wild-type activity
K176A
-
site-directed mutagenesis, significant reduction in catalytic efficiency of enzyme
K176A/E256A
-
double mutant, no activity
K176C
-
wild-type activity
K72A
-
site-directed mutagenesis, wild-type activity
additional information
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
FPLC, anion exchange chromatography, gel filtration
native wild-type enzyme by anion exchange and adsorption chromatography, and gel filtration, recombinant His6-tagged enzyme by nickel affinity chromatography
on nickel-nitrilotriacetic acid resin and by gel filtration, more than 98% pure
anion-exchange chromatography, gel filtration chromatography
-
gel chromatography
-
homogeneity in a yield of 20-60 mg from 3-6 g of cells
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli BL21(DE3)
expression of wild-type and mutant enzymes
gene panE, expression of His6-tagged wild-type and mutant enzymes
KPR expressed using a pRSETA vector in Escherichia coli strain BL21(DE3)C41
expression in Escherichia coli BL21(DE3)
-
expression in Escherichia coli K12
-
overexpression in strain BL21(DE3)
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wilken, D.R.; King, H.L.; Dyar, R.E.
Ketopantoic acid and ketopantoyl lactone reductases. Stereospecificity of transfer of hydrogen from reduced nicotinamide adenine dinucleotide phosphate
J. Biol. Chem.
250
2311-2314
1975
Saccharomyces cerevisiae, Escherichia coli
Manually annotated by BRENDA team
Elischewski, F.; Puhler, A.; Kalinowski, J.
Pantothenate production in Escherichia coli K12 by enhanced expression of the panE gene encoding ketopantoate reductase
J. Biotechnol.
75
135-146
1999
Escherichia coli
Manually annotated by BRENDA team
Frodyma, M.E.; Downs, D.
ApbA, the ketopantoate reductase enzyme of Salmonella typhimurium is required for the synthesis of thiamine via the alternative pyrimidine biosynthetic pathway
J. Biol. Chem.
273
5572-5576
1998
Escherichia coli, Escherichia coli BL21/lambdaDE3, Saccharomyces cerevisiae, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
Manually annotated by BRENDA team
Zheng, R.; Blanchard, J.S.
Kinetic and mechanistic analysis of the E. coli panE-encoded ketopantoate reductase
Biochemistry
39
3708-3717
2000
Escherichia coli
Manually annotated by BRENDA team
Zheng, R.; Blanchard, J.S.
Identification of active site residues in E. coli ketopantoate reductase by mutagenesis and chemical rescue
Biochemistry
39
16244-16251
2000
Escherichia coli
Manually annotated by BRENDA team
Matak-Vinkovic, D.; Vinkovic, M.; Saldanha, S.A.; Ashurst, J.L.; von Delft, F.; Inoue, T.; Miguel, R.N.; Smith, A.G.; Blundell, T.L.; Abell, C.
Crystal structure of Escherichia coli ketopantoate reductase at 1.7 A resolution and insight into the enzyme mechanism
Biochemistry
40
14493-14500
2001
Escherichia coli (P0A9J4), Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
Manually annotated by BRENDA team
Zheng, R.; Blanchard, J.S.
Substrate specificity and kinetic isotope effect analysis of the Eschericia coli ketopantoate reductase
Biochemistry
42
11289-11296
2003
Escherichia coli
Manually annotated by BRENDA team
Ciulli, A.; Abell, C.
Biophysical tools to monitor enzyme-ligand interactions of enzymes involved in vitamin biosynthesis
Biochem. Soc. Trans.
33
767-771
2005
Escherichia coli
Manually annotated by BRENDA team
Lobley, C.M.; Ciulli, A.; Whitney, H.M.; Williams, G.; Smith, A.G.; Abell, C.; Blundell, T.L.
The crystal structure of Escherichia coli ketopantoate reductase with NADP+ bound
Biochemistry
44
8930-8939
2005
Escherichia coli (P0A9J4), Escherichia coli
Manually annotated by BRENDA team
Ciulli, A.; Williams, G.; Smith, A.G.; Blundell, T.L.; Abell, C.
Probing hot spots at protein-ligand binding sites: a fragment-based approach using biophysical methods
J. Med. Chem.
49
4992-5000
2006
Escherichia coli
Manually annotated by BRENDA team
Ciulli, A.; Lobley, C.M.; Tuck, K.L.; Smith, A.G.; Blundell, T.L.; Abell, C.
pH-tuneable binding of 2-phospho-ADP-ribose to ketopantoate reductase: a structural and calorimetric study
Acta Crystallogr. Sect. D
63
171-178
2007
Escherichia coli (P0A9J4), Escherichia coli
Manually annotated by BRENDA team
Ciulli, A.; Chirgadze, D.Y.; Smith, A.G.; Blundell, T.L.; Abell, C.
Crystal structure of Escherichia coli ketopantoate reductase in a ternary complex with NADP+ and pantoate bound: substrate recognition, conformational change, and cooperativity
J. Biol. Chem.
282
8487-8497
2007
Escherichia coli (P0A9J4), Escherichia coli
Manually annotated by BRENDA team
Headey, S.J.; Vom, A.; Simpson, J.S.; Scanlon, M.J.
Backbone assignments of the 34 kDa ketopantoate reductase from E. coli
Biomol. NMR Assign.
2
93-96
2008
Escherichia coli (P0A9J4)
Manually annotated by BRENDA team
Zhang, B.; Zhang, X.M.; Wang, W.; Liu, Z.Q.; Zheng, Y.G.
Metabolic engineering of Escherichia coli for D-pantothenic acid production
Food Chem.
294
267-275
2019
Escherichia coli, Escherichia coli W3110
Manually annotated by BRENDA team