BRENDA - Enzyme Database show
show all sequences of 1.1.1.346

Purification and characterization of 2,5-diketo-D-gluconate reductase from Corynebacterium sp

Miller, J.V.; Estell, D.A.; Lazarus, R.A.; J. Biol. Chem. 262, 9016-9020 (1987)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
Cu2+
strong inhibition at 0.5 mM
Corynebacterium sp.
Fe3+
strong inhibition at 0.5 mM
Corynebacterium sp.
additional information
not inhibited by 2-dehydro-L-gulonate. There is no observed effect on activity by 0.5 mM solutions of Mg2+, Mn2+, Ca2+, or Co2+, 14 mM 2-mercaptoethanol, 1 mM dithiothreitol, or 1 mM EDTA
Corynebacterium sp.
NADP+
competitive inhibition
Corynebacterium sp.
Ni2+
strong inhibition at 0.5 mM
Corynebacterium sp.
Zn2+
strong inhibition at 0.5 mM
Corynebacterium sp.
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.01
-
NADPH
at pH 6.4 and 25°C
Corynebacterium sp.
0.125
-
NADP+
at pH 9.2 and 25°C
Corynebacterium sp.
26
-
2,5-didehydro-D-gluconate
at pH 6.4 and 25°C
Corynebacterium sp.
155
-
5-dehydro-D-fructose
at pH 6.4 and 25°C
Corynebacterium sp.
160
-
dihydroxyacetone
at pH 6.4 and 25°C
Corynebacterium sp.
204
-
2-dehydro-L-gulonate
at pH 9.2 and 25°C
Corynebacterium sp.
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytosol
-
Corynebacterium sp.
5829
-
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
1 * 34000, SDS-PAGE
Corynebacterium sp.
35000
-
gel filtration
Corynebacterium sp.
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Corynebacterium sp.
-
-
-
Corynebacterium sp. ATCC 31090
-
-
-
Purification (Commentary)
Commentary
Organism
DEAE-cellulose column chromatography, Cibacron blueF 3GA affinity column chromatography, and and TSK gel filtration
Corynebacterium sp.
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0052
-
crude extract, at pH 6.4 and 25°C
Corynebacterium sp.
3.38
-
after 650fold purification, at pH 6.4 and 25°C
Corynebacterium sp.
Storage Stability
Storage Stability
Organism
-70°C, 2 mg/ml enzyme in 20 mM Tris-HC1 (pH 7.5), 6 months, the enzyme remains stable
Corynebacterium sp.
4°C, enzyme solution at pH 6.5-7.5, at least 2 months, the enzyme remains stable
Corynebacterium sp.
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,5-didehydro-D-gluconate + NADPH + H+
a reduction of substrate by NADPH is highly preferred
440304
Corynebacterium sp.
2-dehydro-L-gulonate + NADP+
-
-
-
r
2,5-didehydro-D-gluconate + NADPH + H+
a reduction of substrate by NADPH is highly preferred
440304
Corynebacterium sp. ATCC 31090
2-dehydro-L-gulonate + NADP+
-
-
-
r
2-dehydro-L-gulonate + NADP+
-
440304
Corynebacterium sp.
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
r
5-dehydro-D-fructose + NADPH + H+
-
440304
Corynebacterium sp.
? + NADP+
-
-
-
?
dihydroxyacetone + NADPH + H+
-
440304
Corynebacterium sp.
? + NADP+
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
1 * 34000, SDS-PAGE
Corynebacterium sp.
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
8.3
-
2,5-didehydro-D-gluconate
at pH 6.4 and 25°C
Corynebacterium sp.
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.4
-
for the reduction of 2,5-didehydro-D-gluconate
Corynebacterium sp.
9.2
-
for the oxidation of 2-dehydro-L-gulonate
Corynebacterium sp.
pH Range
pH Minimum
pH Maximum
Commentary
Organism
5
8
for the reduction of 2,5-didehydro-D-gluconate
Corynebacterium sp.
Cofactor
Cofactor
Commentary
Organism
Structure
NADPH
dependent on
Corynebacterium sp.
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.026
-
NADP+
at pH 9.2 and 25°C
Corynebacterium sp.
pI Value
Organism
Commentary
pI Value Maximum
pI Value
Corynebacterium sp.
isoelectric focusing
-
4.4
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NADPH
dependent on
Corynebacterium sp.
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
Cu2+
strong inhibition at 0.5 mM
Corynebacterium sp.
Fe3+
strong inhibition at 0.5 mM
Corynebacterium sp.
additional information
not inhibited by 2-dehydro-L-gulonate. There is no observed effect on activity by 0.5 mM solutions of Mg2+, Mn2+, Ca2+, or Co2+, 14 mM 2-mercaptoethanol, 1 mM dithiothreitol, or 1 mM EDTA
Corynebacterium sp.
NADP+
competitive inhibition
Corynebacterium sp.
Ni2+
strong inhibition at 0.5 mM
Corynebacterium sp.
Zn2+
strong inhibition at 0.5 mM
Corynebacterium sp.
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.026
-
NADP+
at pH 9.2 and 25°C
Corynebacterium sp.
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.01
-
NADPH
at pH 6.4 and 25°C
Corynebacterium sp.
0.125
-
NADP+
at pH 9.2 and 25°C
Corynebacterium sp.
26
-
2,5-didehydro-D-gluconate
at pH 6.4 and 25°C
Corynebacterium sp.
155
-
5-dehydro-D-fructose
at pH 6.4 and 25°C
Corynebacterium sp.
160
-
dihydroxyacetone
at pH 6.4 and 25°C
Corynebacterium sp.
204
-
2-dehydro-L-gulonate
at pH 9.2 and 25°C
Corynebacterium sp.
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
cytosol
-
Corynebacterium sp.
5829
-
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
34000
-
1 * 34000, SDS-PAGE
Corynebacterium sp.
35000
-
gel filtration
Corynebacterium sp.
Purification (Commentary) (protein specific)
Commentary
Organism
DEAE-cellulose column chromatography, Cibacron blueF 3GA affinity column chromatography, and and TSK gel filtration
Corynebacterium sp.
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0052
-
crude extract, at pH 6.4 and 25°C
Corynebacterium sp.
3.38
-
after 650fold purification, at pH 6.4 and 25°C
Corynebacterium sp.
Storage Stability (protein specific)
Storage Stability
Organism
-70°C, 2 mg/ml enzyme in 20 mM Tris-HC1 (pH 7.5), 6 months, the enzyme remains stable
Corynebacterium sp.
4°C, enzyme solution at pH 6.5-7.5, at least 2 months, the enzyme remains stable
Corynebacterium sp.
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2,5-didehydro-D-gluconate + NADPH + H+
a reduction of substrate by NADPH is highly preferred
440304
Corynebacterium sp.
2-dehydro-L-gulonate + NADP+
-
-
-
r
2,5-didehydro-D-gluconate + NADPH + H+
a reduction of substrate by NADPH is highly preferred
440304
Corynebacterium sp. ATCC 31090
2-dehydro-L-gulonate + NADP+
-
-
-
r
2-dehydro-L-gulonate + NADP+
-
440304
Corynebacterium sp.
2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
r
5-dehydro-D-fructose + NADPH + H+
-
440304
Corynebacterium sp.
? + NADP+
-
-
-
?
dihydroxyacetone + NADPH + H+
-
440304
Corynebacterium sp.
? + NADP+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 34000, SDS-PAGE
Corynebacterium sp.
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
8.3
-
2,5-didehydro-D-gluconate
at pH 6.4 and 25°C
Corynebacterium sp.
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.4
-
for the reduction of 2,5-didehydro-D-gluconate
Corynebacterium sp.
9.2
-
for the oxidation of 2-dehydro-L-gulonate
Corynebacterium sp.
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
5
8
for the reduction of 2,5-didehydro-D-gluconate
Corynebacterium sp.
pI Value (protein specific)
Organism
Commentary
pI Value Maximum
pI Value
Corynebacterium sp.
isoelectric focusing
-
4.4
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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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
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-
741575
Kaswurm
Evaluation of the food grade ...
Corynebacterium glutamicum, Corynebacterium glutamicum DSM 20301
AMB Express
3
7-17
2013
-
1
1
-
1
-
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2
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2
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6
-
2
1
1
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2
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1
1
2
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1
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2
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6
-
2
1
1
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-
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1
-
-
-
-
2
2
-
-
-
657308
Sanli
Structural alteration of cofac ...
Corynebacterium sp.
Protein Sci.
13
504-512
2004
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1
1
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1
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1
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-
-
-
-
-
-
-
-
-
-
-
440310
Banta
Alteration of the specificity ...
Corynebacterium sp.
Protein Eng.
15
131-140
2002
-
-
1
-
40
-
-
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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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1
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1
1
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40
-
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1
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1
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2
1
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-
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-
-
-
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-
440311
Banta
Optimizing an artificial metab ...
Corynebacterium sp.
Biochemistry
41
6226-6236
2002
-
-
-
-
5
-
-
-
-
-
-
-
-
1
-
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1
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5
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1
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1
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5
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-
1
-
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-
-
5
-
-
-
-
-
-
-
-
-
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440312
Habrych
Purification and identificatio ...
Escherichia coli, Escherichia coli BL21-(DE3)
Biotechnol. Prog.
18
257-261
2002
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-
-
-
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1
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1
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2
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1
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1
1
5
1
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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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1
-
-
-
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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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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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-
-
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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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3
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1
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2
1
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1
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-
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-
-
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286278
Yum
Purification and characterizat ...
Brevibacterium ketosoreductum, Brevibacterium ketosoreductum ATCC 21914
Biosci. Biotechnol. Biochem.
62
154-156
1998
-
-
-
-
-
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-
2
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2
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2
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1
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2
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5
1
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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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1
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2
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5
1
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-
1
-
1
1
-
-
-
-
-
-
440305
Khurana
Crystal structure of 2,5-diket ...
Corynebacterium sp.
Proc. Natl. Acad. Sci. USA
95
6768-6773
1998
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-
1
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
1
1
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1
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1
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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
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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
1
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1
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1
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-
440303
Sonoyama
-
Purification and properties of ...
Corynebacterium sp., Corynebacterium sp. SHS 0007
J. Ferment. Technol.
65
311-317
1987
-
-
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4
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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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2
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1
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1
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4
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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
2
-
-
-
-
-
-
440304
Miller
Purification and characterizat ...
Corynebacterium sp., Corynebacterium sp. ATCC 31090
J. Biol. Chem.
262
9016-9020
1987
-
-
-
-
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6
6
1
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2
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-
2
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1
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2
2
5
1
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1
2
1
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1
1
1
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1
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6
1
6
1
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2
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1
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2
2
5
1
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1
2
1
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1
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