BRENDA - Enzyme Database show
show all sequences of 1.1.1.346

Purification and identification of an Escherichia coli beta-keto ester reductase as 2,5-diketo-D-gluconate reductase YqhE

Habrych, M.; Rodriguez, S.; Stewart, J.D.; Biotechnol. Prog. 18, 257-261 (2002)

Data extracted from this reference:

KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1
-
ethyl 2-methylacetoacetate
at pH 7.0 and 30C
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
31800
-
x * 31800, estimated from SDS-PAGE
Escherichia coli
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Escherichia coli BL21-(DE3)
-
-
-
Purification (Commentary)
Commentary
Organism
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, Matrex Red-A column chromatography, Sephacryl S-200 gel filtration, and hypatite C cholumn chromatography
Escherichia coli
Specific Activity [micromol/min/mg]
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
6.3
-
purified enzyme, at pH 7.0 and 30C
Escherichia coli
Storage Stability
Storage Stability
Organism
4C, potassium buffer, several weeks, no loss of activity
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ethyl 2-acetylpent-4-enoate + NADPH + H+
250% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (2R)-2-[(1S)-1-hydroxyethyl]pent-4-enoate + NADP+
-
-
-
?
ethyl 2-ethyl-3-oxobutanoate + NADPH + H+
120% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (2R,3S)-2-ethyl-3-hydroxybutanoate + NADP+
-
-
-
?
ethyl 2-methylacetoacetate + NADH + H+
7% activity with NADH compared to NADPH
440312
Escherichia coli
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NAD+
-
-
-
?
ethyl 2-methylacetoacetate + NADPH + H+
-
440312
Escherichia coli
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
-
-
?
ethyl acetoacetate + NADPH + H+
53% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (3S)-3-hydroxybutanoate + NADP+
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 31800, estimated from SDS-PAGE
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.6
-
ethyl 2-methylacetoacetate
at pH 7.0 and 30C
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
dependent on
Escherichia coli
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Escherichia coli
isoelectric focusing
-
5.9
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
dependent on
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1
-
ethyl 2-methylacetoacetate
at pH 7.0 and 30C
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
31800
-
x * 31800, estimated from SDS-PAGE
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
Organism
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, Matrex Red-A column chromatography, Sephacryl S-200 gel filtration, and hypatite C cholumn chromatography
Escherichia coli
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
6.3
-
purified enzyme, at pH 7.0 and 30C
Escherichia coli
Storage Stability (protein specific)
Storage Stability
Organism
4C, potassium buffer, several weeks, no loss of activity
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ethyl 2-acetylpent-4-enoate + NADPH + H+
250% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (2R)-2-[(1S)-1-hydroxyethyl]pent-4-enoate + NADP+
-
-
-
?
ethyl 2-ethyl-3-oxobutanoate + NADPH + H+
120% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (2R,3S)-2-ethyl-3-hydroxybutanoate + NADP+
-
-
-
?
ethyl 2-methylacetoacetate + NADH + H+
7% activity with NADH compared to NADPH
440312
Escherichia coli
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NAD+
-
-
-
?
ethyl 2-methylacetoacetate + NADPH + H+
-
440312
Escherichia coli
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
-
-
?
ethyl acetoacetate + NADPH + H+
53% activity compared to ethyl 2-methylacetoacetate
440312
Escherichia coli
ethyl (3S)-3-hydroxybutanoate + NADP+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 31800, estimated from SDS-PAGE
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.6
-
ethyl 2-methylacetoacetate
at pH 7.0 and 30C
Escherichia coli
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Escherichia coli
isoelectric focusing
-
5.9
General Information
General Information
Commentary
Organism
metabolism
the enzyme reduces 2,5-didehydro-D-gluconate, a key step in the microbial synthesis of vitamin C
Escherichia coli
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the enzyme reduces 2,5-didehydro-D-gluconate, a key step in the microbial synthesis of vitamin C
Escherichia coli
Other publictions for EC 1.1.1.346
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)
742259
Chen
Cloning, expression and chara ...
Comamonas testosteroni, Comamonas testosteroni ATCC 11996
Chem. Biol. Interact.
234
229-235
2015
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1
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1
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4
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2
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1
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4
1
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1
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1
1
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1
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4
-
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1
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4
1
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-
-
-
-
-
-
1
3
3
1
-
-
741575
Kaswurm
Evaluation of the food grade ...
Corynebacterium glutamicum, Corynebacterium glutamicum DSM 20301
AMB Express
3
7-17
2013
-
1
1
-
1
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
6
-
2
1
1
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1
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2
-
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1
1
2
-
1
-
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2
-
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-
-
-
6
-
2
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
657308
Sanli
Structural alteration of cofac ...
Corynebacterium sp.
Protein Sci.
13
504-512
2004
-
-
-
1
1
-
-
-
-
-
-
-
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1
-
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1
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1
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1
1
1
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
440310
Banta
Alteration of the specificity ...
Corynebacterium sp.
Protein Eng.
15
131-140
2002
-
-
1
-
40
-
-
-
-
-
1
-
-
2
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
40
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
440311
Banta
Optimizing an artificial metab ...
Corynebacterium sp.
Biochemistry
41
6226-6236
2002
-
-
-
-
5
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
5
-
-
-
1
-
-
-
-
-
-
1
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
440312
Habrych
Purification and identificatio ...
Escherichia coli, Escherichia coli BL21-(DE3)
Biotechnol. Prog.
18
257-261
2002
-
-
-
-
-
-
-
1
-
-
1
-
-
2
-
-
1
-
-
-
1
1
5
1
-
-
-
1
-
-
-
1
-
1
-
-
-
-
1
-
-
-
-
-
-
1
-
-
1
-
-
-
-
1
-
-
1
1
5
1
-
-
-
1
-
-
-
1
-
1
1
-
-
-
440307
Khurana
Molecular modeling of substrat ...
Corynebacterium sp.
Proteins
39
68-75
2000
-
-
-
-
2
-
-
-
-
-
-
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-
1
-
-
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-
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1
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1
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1
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2
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-
1
-
-
-
-
-
-
-
-
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-
-
-
-
440306
Yum
Identification of the yqhE and ...
Escherichia coli
Appl. Environ. Microbiol.
65
3341-3346
1999
-
-
-
-
-
-
-
-
-
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3
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8
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1
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-
2
1
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-
-
1
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-
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3
-
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1
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-
2
1
-
-
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-
1
-
-
-
-
-
-
-
-
-
286278
Yum
Purification and characterizat ...
Brevibacterium ketosoreductum, Brevibacterium ketosoreductum ATCC 21914
Biosci. Biotechnol. Biochem.
62
154-156
1998
-
-
-
-
-
-
-
2
-
-
2
-
-
2
-
-
1
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-
-
2
-
5
1
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-
-
1
-
1
1
-
1
-
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-
-
1
-
-
-
-
-
-
2
-
-
2
-
-
-
-
1
-
-
2
-
5
1
-
-
-
-
1
-
1
1
-
-
-
-
-
-
440305
Khurana
Crystal structure of 2,5-diket ...
Corynebacterium sp.
Proc. Natl. Acad. Sci. USA
95
6768-6773
1998
-
-
1
1
-
-
-
-
-
-
-
-
-
3
-
-
1
-
-
-
-
-
1
-
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-
1
-
-
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1
1
1
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-
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1
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1
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-
-
-
-
-
-
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-
721898
Maremonti
-
Characterisation of 2,5-diketo ...
Corynebacterium sp.
Biotechnol. Lett.
18
845-850
1996
-
-
-
-
-
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-
1
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2
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1
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1
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2
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1
1
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1
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1
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1
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2
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1
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2
-
1
1
-
-
1
-
-
-
-
1
-
-
-
-
-
-
440303
Sonoyama
-
Purification and properties of ...
Corynebacterium sp., Corynebacterium sp. SHS 0007
J. Ferment. Technol.
65
311-317
1987
-
-
-
-
-
-
-
4
-
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2
-
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2
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1
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5
1
1
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1
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2
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1
1
-
2
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1
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4
-
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2
-
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1
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-
-
-
5
1
1
-
1
-
2
-
1
2
-
-
-
-
-
-
440304
Miller
Purification and characterizat ...
Corynebacterium sp., Corynebacterium sp. ATCC 31090
J. Biol. Chem.
262
9016-9020
1987
-
-
-
-
-
-
6
6
1
-
2
-
-
2
-
-
1
-
-
-
2
2
5
1
-
-
-
1
2
1
-
1
1
1
-
-
-
-
1
-
-
-
-
6
1
6
1
-
2
-
-
-
-
1
-
-
2
2
5
1
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-
-
1
2
1
-
1
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-
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-
-