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Literature summary extracted from

  • Miller, J.V.; Estell, D.A.; Lazarus, R.A.
    Purification and characterization of 2,5-diketo-D-gluconate reductase from Corynebacterium sp (1987), J. Biol. Chem., 262, 9016-9020.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.274 Cu2+ 0.5 mM, strong inhibition Corynebacterium sp.
1.1.1.274 Fe3+ 0.5 mM, strong inhibition Corynebacterium sp.
1.1.1.274 NADP+ competitive product inhibition Corynebacterium sp.
1.1.1.274 Ni2+ 0.5 mM, strong inhibition Corynebacterium sp.
1.1.1.274 Zn2+ 0.5 mM, strong inhibition Corynebacterium sp.
1.1.1.346 Cu2+ strong inhibition at 0.5 mM Corynebacterium sp.
1.1.1.346 Fe3+ strong inhibition at 0.5 mM Corynebacterium sp.
1.1.1.346 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.
1.1.1.346 NADP+ competitive inhibition Corynebacterium sp.
1.1.1.346 Ni2+ strong inhibition at 0.5 mM Corynebacterium sp.
1.1.1.346 Zn2+ strong inhibition at 0.5 mM Corynebacterium sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.274 0.01
-
NADPH
-
Corynebacterium sp.
1.1.1.274 2.2
-
glycerol
-
Corynebacterium sp.
1.1.1.274 26
-
2,5-didehydro-D-gluconate
-
Corynebacterium sp.
1.1.1.274 155
-
5-keto-D-fructose
-
Corynebacterium sp.
1.1.1.274 160
-
dihydroxyacetone
-
Corynebacterium sp.
1.1.1.346 0.01
-
NADPH at pH 6.4 and 25°C Corynebacterium sp.
1.1.1.346 0.125
-
NADP+ at pH 9.2 and 25°C Corynebacterium sp.
1.1.1.346 26
-
2,5-didehydro-D-gluconate at pH 6.4 and 25°C Corynebacterium sp.
1.1.1.346 155
-
5-dehydro-D-fructose at pH 6.4 and 25°C Corynebacterium sp.
1.1.1.346 160
-
dihydroxyacetone at pH 6.4 and 25°C Corynebacterium sp.
1.1.1.346 204
-
2-dehydro-L-gulonate at pH 9.2 and 25°C Corynebacterium sp.

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.1.1.346 cytosol
-
Corynebacterium sp. 5829
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.274 29992
-
1 * 29992, deduced from nucleotide sequence Corynebacterium sp.
1.1.1.274 34000
-
1 * 34000, SDS-PAGE Corynebacterium sp.
1.1.1.274 35000
-
gel filtration Corynebacterium sp.
1.1.1.346 34000
-
1 * 34000, SDS-PAGE Corynebacterium sp.
1.1.1.346 35000
-
gel filtration Corynebacterium sp.

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.274 Corynebacterium sp.
-
-
-
1.1.1.346 Corynebacterium sp.
-
-
-
1.1.1.346 Corynebacterium sp. ATCC 31090
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.274 DEAE-cellulose, Cibacron blue F3GA agarose, gel filtration Corynebacterium sp.
1.1.1.346 DEAE-cellulose column chromatography, Cibacron blueF 3GA affinity column chromatography, and and TSK gel filtration Corynebacterium sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.274 3.38
-
-
Corynebacterium sp.
1.1.1.346 0.0052
-
crude extract, at pH 6.4 and 25°C Corynebacterium sp.
1.1.1.346 3.38
-
after 650fold purification, at pH 6.4 and 25°C Corynebacterium sp.

Storage Stability

EC Number Storage Stability Organism
1.1.1.274 -70°C, 20 mM Tris-HCl, pH 7.5, over 6 months, no loss of activity Corynebacterium sp.
1.1.1.274 4°C, pH 6.5-7.5, at least 2 months, no loss of activity Corynebacterium sp.
1.1.1.346 -70°C, 2 mg/ml enzyme in 20 mM Tris-HC1 (pH 7.5), 6 months, the enzyme remains stable Corynebacterium sp.
1.1.1.346 4°C, enzyme solution at pH 6.5-7.5, at least 2 months, the enzyme remains stable Corynebacterium sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.274 2,5-didehydro-D-gluconate + NADH 170fold lower reduction rate with NADH compared to NADPH Corynebacterium sp. 2-keto-L-gulonate + NAD+
-
?
1.1.1.274 2,5-didehydro-D-gluconate + NADH 170fold lower reduction rate with NADH compared to NADPH Corynebacterium sp. ATCC 31090 2-keto-L-gulonate + NAD+
-
?
1.1.1.274 2,5-didehydro-D-gluconate + NADPH stereospecific reduction to 2-keto-L-gulonate, no activity with D-fructose, L-sorbose, 5-keto-D-gluconate, 2-keto-L-gulonate, 2-keto-D-gluconate, pyruvate or hydroxypyruvate Corynebacterium sp. 2-keto-L-gulonate + NADP+
-
r
1.1.1.274 2,5-didehydro-D-gluconate + NADPH stereospecific reduction to 2-keto-L-gulonate, no activity with D-fructose, L-sorbose, 5-keto-D-gluconate, 2-keto-L-gulonate, 2-keto-D-gluconate, pyruvate or hydroxypyruvate Corynebacterium sp. ATCC 31090 2-keto-L-gulonate + NADP+
-
r
1.1.1.274 5-keto-D-fructose + NADPH
-
Corynebacterium sp. L-sorbose + NADP+
-
?
1.1.1.274 5-keto-D-fructose + NADPH
-
Corynebacterium sp. ATCC 31090 L-sorbose + NADP+
-
?
1.1.1.274 dihydroxyacetone + NADPH
-
Corynebacterium sp. glycerol + NADP+
-
?
1.1.1.274 dihydroxyacetone + NADPH
-
Corynebacterium sp. ATCC 31090 glycerol + NADP+
-
?
1.1.1.346 2,5-didehydro-D-gluconate + NADPH + H+ a reduction of substrate by NADPH is highly preferred Corynebacterium sp. 2-dehydro-L-gulonate + NADP+
-
r
1.1.1.346 2,5-didehydro-D-gluconate + NADPH + H+ a reduction of substrate by NADPH is highly preferred Corynebacterium sp. ATCC 31090 2-dehydro-L-gulonate + NADP+
-
r
1.1.1.346 2-dehydro-L-gulonate + NADP+
-
Corynebacterium sp. 2,5-didehydro-D-gluconate + NADPH + H+
-
r
1.1.1.346 5-dehydro-D-fructose + NADPH + H+
-
Corynebacterium sp. ? + NADP+
-
?
1.1.1.346 dihydroxyacetone + NADPH + H+
-
Corynebacterium sp. ? + NADP+
-
?

Subunits

EC Number Subunits Comment Organism
1.1.1.274 monomer 1 * 34000, SDS-PAGE Corynebacterium sp.
1.1.1.274 monomer 1 * 29992, deduced from nucleotide sequence Corynebacterium sp.
1.1.1.346 monomer 1 * 34000, SDS-PAGE Corynebacterium sp.

Synonyms

EC Number Synonyms Comment Organism
1.1.1.346 2,5-diketo-D-gluconate reductase
-
Corynebacterium sp.
1.1.1.346 2,5-DKG reductase
-
Corynebacterium sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.274 8.33
-
2,5-didehydro-D-gluconate
-
Corynebacterium sp.
1.1.1.346 8.3
-
2,5-didehydro-D-gluconate at pH 6.4 and 25°C Corynebacterium sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.274 6.4
-
reduction of 2,5-didehydro-D-gluconate Corynebacterium sp.
1.1.1.274 9.2
-
oxidation of 2-keto-L-gulonate Corynebacterium sp.
1.1.1.346 6.4
-
for the reduction of 2,5-didehydro-D-gluconate Corynebacterium sp.
1.1.1.346 9.2
-
for the oxidation of 2-dehydro-L-gulonate Corynebacterium sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.274 4.5 9.3 reduction of 2,5-didehydro-D-gluconate Corynebacterium sp.
1.1.1.274 8 10.3 oxidation of 2-keto-L-gulonate Corynebacterium sp.
1.1.1.346 5 8 for the reduction of 2,5-didehydro-D-gluconate Corynebacterium sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.274 NADP+
-
Corynebacterium sp.
1.1.1.274 NADPH
-
Corynebacterium sp.
1.1.1.346 NADPH dependent on Corynebacterium sp.

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.274 0.026
-
NADP+
-
Corynebacterium sp.
1.1.1.346 0.026
-
NADP+ at pH 9.2 and 25°C Corynebacterium sp.

pI Value

EC Number Organism Comment pI Value Maximum pI Value
1.1.1.346 Corynebacterium sp. isoelectric focusing
-
4.4