BRENDA - Enzyme Database show
show all sequences of 1.1.1.169

Crystal structure of Escherichia coli ketopantoate reductase in a ternary complex with NADP+ and pantoate bound: substrate recognition, conformational change, and cooperativity

Ciulli, A.; Chirgadze, D.Y.; Smith, A.G.; Blundell, T.L.; Abell, C.; J. Biol. Chem. 282, 8487-8497 (2007)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of wild-type and mutant enzymes
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
purified recombinant His6-tagged enzyme in complex with NADP+ and pantoate, hanging drop vapor-diffusion technique, 20C, 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
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
E256A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
K176A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
K72A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
N98A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
R31A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
S244A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of recombinant wild-type and mutant enzymes, overview
Escherichia coli
0.007
-
NADPH
pH 7.5, 27C, wild-type enzyme
Escherichia coli
0.03
-
2-dehydropantoate
pH 7.5, 27C, wild-type enzyme
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-dehydropantoate + NADPH
Escherichia coli
an essential step for the biosynthesis of pantothenate, i.e. vitamin B5
(R)-pantoate + NADP+
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P0A9J4
gene panE
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-dehydropantoate + NADPH
an essential step for the biosynthesis of pantothenate, i.e. vitamin B5
687675
Escherichia coli
(R)-pantoate + NADP+
-
-
-
r
2-dehydropantoate + NADPH
substrate and product binding structures, hinge bending between the N- and C-terminal domains is observed, which triggers the switch of the essential Lys176 to form a key hydrogen bond with the C2 hydroxyl of pantoate, pantoate forms additional interactions with conserved residues Ser244, Asn98, and Asn180 and with two conservatively varied residues, Asn194 and Asn241, overview
687675
Escherichia coli
(R)-pantoate + NADP+
-
-
-
r
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
27
-
assay at
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
25
-
2-dehydropantoate
pH 7.5, 27C, wild-type enzyme
Escherichia coli
25
-
NADPH
pH 7.5, 27C, wild-type enzyme
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of wild-type and mutant enzymes
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant His6-tagged enzyme in complex with NADP+ and pantoate, hanging drop vapor-diffusion technique, 20C, 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
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
E256A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
K176A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
K72A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
N98A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
R31A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
S244A
site-directed mutagenesis, altered steady-state kinetics of the mutant compared to the wild-type enzyme, overview
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of recombinant wild-type and mutant enzymes, overview
Escherichia coli
0.007
-
NADPH
pH 7.5, 27C, wild-type enzyme
Escherichia coli
0.03
-
2-dehydropantoate
pH 7.5, 27C, wild-type enzyme
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2-dehydropantoate + NADPH
Escherichia coli
an essential step for the biosynthesis of pantothenate, i.e. vitamin B5
(R)-pantoate + NADP+
-
-
r
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-dehydropantoate + NADPH
an essential step for the biosynthesis of pantothenate, i.e. vitamin B5
687675
Escherichia coli
(R)-pantoate + NADP+
-
-
-
r
2-dehydropantoate + NADPH
substrate and product binding structures, hinge bending between the N- and C-terminal domains is observed, which triggers the switch of the essential Lys176 to form a key hydrogen bond with the C2 hydroxyl of pantoate, pantoate forms additional interactions with conserved residues Ser244, Asn98, and Asn180 and with two conservatively varied residues, Asn194 and Asn241, overview
687675
Escherichia coli
(R)-pantoate + NADP+
-
-
-
r
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
27
-
assay at
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
25
-
2-dehydropantoate
pH 7.5, 27C, wild-type enzyme
Escherichia coli
25
-
NADPH
pH 7.5, 27C, wild-type enzyme
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Escherichia coli
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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-
-
-
-
-
-
-
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-
12
-
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-
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-
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1
1
-
-
-
736827
Tomita
Identification and characteriz ...
Thermococcus kodakarensis, Thermococcus kodakarensis KUW1
Mol. Microbiol.
90
307-321
2013
-
-
1
-
1
-
1
7
-
-
2
4
-
8
-
-
1
-
-
-
-
-
13
1
1
1
1
6
1
-
-
4
1
-
-
-
-
1
4
-
1
-
-
1
1
7
-
-
2
4
-
-
-
1
-
-
-
-
13
1
1
1
1
6
1
-
-
-
-
2
2
-
6
6
696603
Headey
Backbone assignments of the 34 ...
Escherichia coli
Biomol. NMR Assign.
2
93-96
2008
-
-
1
-
-
-
-
-
-
-
1
-
-
1
-
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1
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1
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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
-
-
-
-
-
-
-
1
-
2
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3
-
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-
1
-
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-
1
1
-
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-
-
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-
1
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
687675
Ciulli
Crystal structure of Escherich ...
Escherichia coli
J. Biol. Chem.
282
8487-8497
2007
-
-
1
1
6
-
-
3
-
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-
1
-
2
-
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-
-
-
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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
-
2
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-
1
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-
2
-
-
-
-
-
1
-
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3
6
-
-
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-
3
-
-
-
-
7
6
1
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
667561
Ciulli
Biophysical tools to monitor e ...
Escherichia coli
Biochem. Soc. Trans.
33
767-771
2005
-
-
-
-
-
-
1
-
-
-
-
1
-
1
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2
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2
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2
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1
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1
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-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
667687
Lobley
The crystal structure of Esche ...
Escherichia coli
Biochemistry
44
8930-8939
2005
-
-
1
1
7
-
-
1
-
-
-
1
-
2
-
-
1
1
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
1
7
-
-
-
-
1
-
-
-
1
-
-
-
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
654677
Zheng
Substrate specificity and kine ...
Escherichia coli
Biochemistry
42
11289-11296
2003
-
-
1
-
-
-
-
8
-
-
-
1
-
2
-
-
-
-
-
-
-
-
7
-
1
-
-
-
1
1
-
4
-
-
-
-
-
1
4
-
-
-
-
-
-
8
-
-
-
1
-
-
-
-
-
-
-
-
7
-
1
-
-
-
1
1
-
-
-
-
-
-
-
-
656330
Merkamm
Ketopantoate reductase activit ...
Corynebacterium glutamicum
J. Biotechnol.
104
253-260
2003
-
-
1
-
-
-
-
1
-
-
-
1
-
3
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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
-
-
-
2
-
4
3
-
-
-
2
-
-
-
3
-
-
-
-
-
1
3
1
1
-
-
-
-
4
-
2
4
2
-
-
1
1
-
-
2
-
4
3
-
-
-
2
-
-
-
-
-
-
-
-
-
-
286108
Zheng
Kinetic and mechanistic analys ...
Escherichia coli
Biochemistry
39
3708-3717
2000
-
-
1
-
-
-
2
4
-
1
4
1
-
2
-
-
1
-
-
-
1
-
2
1
-
-
-
-
-
-
-
2
-
-
-
-
-
1
2
-
-
-
-
2
-
4
-
1
4
1
-
-
-
1
-
-
1
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
286109
Zheng
Identification of active site ...
Escherichia coli
Biochemistry
39
16244-16251
2000
5
-
1
-
10
-
-
12
-
1
1
-
-
2
-
-
1
-
-
-
1
-
2
1
-
-
-
-
1
-
-
2
-
-
-
5
-
1
2
-
10
-
-
-
-
12
-
1
1
-
-
-
-
1
-
-
1
-
2
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
286106
Elischewski
Pantothenate production in Esc ...
Escherichia coli
J. Biotechnol.
75
135-146
1999
-
-
1
-
-
-
-
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1
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3
-
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1
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1
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1
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1
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-
-
-
-
-
-
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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
-
-
1
-
-
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-
2
-
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1
3
-
10
-
-
1
-
-
-
6
-
12
2
1
-
-
-
1
-
-
8
-
-
-
-
-
1
8
-
-
-
-
-
-
2
-
-
1
3
-
-
-
1
-
-
6
-
12
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
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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-
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4
-
-
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-
-
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4
-
2
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3
-
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3
-
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-
-
-
4
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
286103
Shimizu
Ketopantoic acid reductase of ...
Saccharomyces cerevisiae, Stenotrophomonas maltophilia, Stenotrophomonas maltophilia 845
J. Biol. Chem.
263
12077-12084
1988
-
-
-
1
-
-
10
4
-
1
4
-
-
6
-
-
1
-
-
-
2
1
6
1
1
1
3
-
2
-
2
4
-
-
-
-
-
-
4
1
-
-
-
10
-
4
-
1
4
-
-
-
-
1
-
-
2
1
6
1
1
1
3
-
2
-
2
-
-
-
-
-
-
-
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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-
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-
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2
-
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-
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-
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2
-
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-
-
2
-
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-
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286104
King
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Separation and preliminary stu ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae NRRL Y-2034
J. Biol. Chem.
247
4096-4098
1972
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