BRENDA - Enzyme Database show
show all sequences of 1.1.1.169

The crystal structure of Escherichia coli ketopantoate reductase with NADP+ bound

Lobley, C.M.; Ciulli, A.; Whitney, H.M.; Williams, G.; Smith, A.G.; Abell, C.; Blundell, T.L.; Biochemistry 44, 8930-8939 (2005)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene panE, expression of His6-tagged wild-type and mutant enzymes
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
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
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
D248A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
E210A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
E256A
site-directed mutagenesis, nearly inactive mutant, 2600fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
K176A
site-directed mutagenesis, nearly inactive mutant, 78000fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
K72A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
N98A
site-directed mutagenesis, nearly inactive mutant, 4000fold reduced catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
S244A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics and thermodynamics, wild-type enzyme, overview
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ketopantoate + NADPH
Escherichia coli
the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA
pantoate + NADP+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P0A9J4
gene panE
-
Purification (Commentary)
Commentary
Organism
native wild-type enzyme by anion exchange and adsorption chromatography, and gel filtration, recombinant His6-tagged enzyme by nickel affinity chromatography
Escherichia coli
Reaction
Reaction
Commentary
Organism
(R)-pantoate + NADP+ = 2-dehydropantoate + NADPH + H+
molecular catalytic mechanism, substrate and cofactor binding, Asn98, Glu256, and Lys176 are essential, overview
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ketopantoate + NADPH
the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA
667687
Escherichia coli
pantoate + NADP+
-
-
-
r
ketopantoate + NADPH
substrate binding structure and thermodynamics
667687
Escherichia coli
pantoate + NADP+
-
-
-
r
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
cofactor binding structure and thermodynamics, the cofactor is bound in the active site cleft between the N-terminal Rossmann-fold domain and the C-terminal alpha-helical domain
Escherichia coli
NADPH
cofactor binding structure and thermodynamics
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
gene panE, expression of His6-tagged wild-type and mutant enzymes
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
cofactor binding structure and thermodynamics, the cofactor is bound in the active site cleft between the N-terminal Rossmann-fold domain and the C-terminal alpha-helical domain
Escherichia coli
NADPH
cofactor binding structure and thermodynamics
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
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
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D248A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
E210A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
E256A
site-directed mutagenesis, nearly inactive mutant, 2600fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
K176A
site-directed mutagenesis, nearly inactive mutant, 78000fold decreased catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
K72A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
N98A
site-directed mutagenesis, nearly inactive mutant, 4000fold reduced catalytic efficiency, no complementation of a panE knockout mutant strain
Escherichia coli
S244A
site-directed mutagenesis, unaltered activity compared to the wild-type enzyme, functional complementation of a panE knockout mutant strain
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics and thermodynamics, wild-type enzyme, overview
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ketopantoate + NADPH
Escherichia coli
the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA
pantoate + NADP+
-
-
r
Purification (Commentary) (protein specific)
Commentary
Organism
native wild-type enzyme by anion exchange and adsorption chromatography, and gel filtration, recombinant His6-tagged enzyme by nickel affinity chromatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ketopantoate + NADPH
the enzyme is involved in pantothenate, i.e. vitamin B5, biosynthesis, which is a precursor for CoA
667687
Escherichia coli
pantoate + NADP+
-
-
-
r
ketopantoate + NADPH
substrate binding structure and thermodynamics
667687
Escherichia coli
pantoate + NADP+
-
-
-
r
Other publictions for EC 1.1.1.169
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
725293
Miller
PanG, a new ketopantoate reduc ...
Francisella tularensis subsp. novicida
J. Bacteriol.
195
965-976
2013
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12
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1
1
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736827
Tomita
Identification and characteriz ...
Thermococcus kodakarensis, Thermococcus kodakarensis KUW1
Mol. Microbiol.
90
307-321
2013
-
-
1
-
1
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1
7
-
-
2
4
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8
-
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1
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13
1
1
1
1
6
1
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4
1
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1
4
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1
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1
1
7
-
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2
4
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1
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13
1
1
1
1
6
1
-
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-
2
2
-
6
6
696603
Headey
Backbone assignments of the 34 ...
Escherichia coli
Biomol. NMR Assign.
2
93-96
2008
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1
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1
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1
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1
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684140
Ciulli
pH-tuneable binding of 2-phosp ...
Escherichia coli
Acta Crystallogr. Sect. D
63
171-178
2007
-
-
-
1
-
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-
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-
1
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2
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3
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1
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1
1
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1
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3
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687675
Ciulli
Crystal structure of Escherich ...
Escherichia coli
J. Biol. Chem.
282
8487-8497
2007
-
-
1
1
6
-
-
3
-
-
-
1
-
2
-
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-
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-
2
-
1
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2
1
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2
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1
2
1
6
-
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3
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1
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2
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1
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2
1
-
-
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669806
Ciulli
Probing hot spots at protein-l ...
Escherichia coli
J. Med. Chem.
49
4992-5000
2006
-
-
-
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7
1
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1
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2
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1
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2
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1
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3
6
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3
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7
6
1
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1
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2
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1
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667561
Ciulli
Biophysical tools to monitor e ...
Escherichia coli
Biochem. Soc. Trans.
33
767-771
2005
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1
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1
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1
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667687
Lobley
The crystal structure of Esche ...
Escherichia coli
Biochemistry
44
8930-8939
2005
-
-
1
1
7
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1
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1
-
2
-
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1
1
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2
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2
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1
2
1
7
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1
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1
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1
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2
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654677
Zheng
Substrate specificity and kine ...
Escherichia coli
Biochemistry
42
11289-11296
2003
-
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1
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8
-
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1
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2
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7
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1
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1
1
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4
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1
4
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8
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1
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7
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1
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1
1
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-
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-
656330
Merkamm
Ketopantoate reductase activit ...
Corynebacterium glutamicum
J. Biotechnol.
104
253-260
2003
-
-
1
-
-
-
-
1
-
-
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1
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3
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2
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2
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1
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1
1
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1
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1
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2
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2
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-
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286110
Matak-Vinkovic
Crystal structure of Escherich ...
Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
Biochemistry
40
14493-14500
2001
-
-
1
1
1
-
-
4
-
2
4
2
-
7
-
1
1
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-
-
2
-
4
3
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2
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3
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1
3
1
1
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4
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2
4
2
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1
1
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2
-
4
3
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-
2
-
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286108
Zheng
Kinetic and mechanistic analys ...
Escherichia coli
Biochemistry
39
3708-3717
2000
-
-
1
-
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-
2
4
-
1
4
1
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2
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1
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1
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2
1
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2
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1
2
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2
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4
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1
4
1
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1
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1
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2
1
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286109
Zheng
Identification of active site ...
Escherichia coli
Biochemistry
39
16244-16251
2000
5
-
1
-
10
-
-
12
-
1
1
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-
2
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1
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1
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2
1
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1
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2
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5
-
1
2
-
10
-
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12
-
1
1
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1
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1
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2
1
-
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1
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286106
Elischewski
Pantothenate production in Esc ...
Escherichia coli
J. Biotechnol.
75
135-146
1999
-
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1
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1
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3
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1
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286107
Frodyma
ApbA, the ketopantoate reducta ...
Escherichia coli, Escherichia coli BL21/lambdaDE3, Saccharomyces cerevisiae, Salmonella enterica subsp. enterica serovar Typhimurium, Stenotrophomonas maltophilia 845, Stenotrophomonas maltophilia
J. Biol. Chem.
273
5572-5576
1998
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1
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2
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1
3
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10
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1
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6
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12
2
1
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1
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8
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8
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1
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6
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12
2
1
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1
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286102
Kataoka
-
Novel enzymic production of D- ...
Agrobacterium sp., Agrobacterium tumefaciens, Pseudomonas putida, Stenotrophomonas maltophilia
Agric. Biol. Chem.
54
177-182
1990
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4
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2
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286103
Shimizu
Ketopantoic acid reductase of ...
Saccharomyces cerevisiae, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
J. Biol. Chem.
263
12077-12084
1988
-
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1
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10
4
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1
4
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6
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1
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2
1
6
1
1
1
3
-
2
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2
4
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4
1
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10
-
4
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1
4
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1
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2
1
6
1
1
1
3
-
2
-
2
-
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-
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286105
Wilken
Ketopantoic acid and ketopanto ...
Escherichia coli, Saccharomyces cerevisiae
J. Biol. Chem.
250
2311-2314
1975
-
-
-
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2
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2
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2
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2
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2
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2
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2
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286104
King
-
Separation and preliminary stu ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae NRRL Y-2034
J. Biol. Chem.
247
4096-4098
1972
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2
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1
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4
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1
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2
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1
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2
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4
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1
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