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Literature summary extracted from

  • Banta, S.; Swanson, B.A.; Wu, S.; Jarnagin, A.; Anderson, S.
    Optimizing an artificial metabolic pathway: engineering the cofactor specificity of Corynebacterium 2,5-diketo-D-gluconic acid reductase for use in vitamin C biosynthesis (2002), Biochemistry, 41, 6226-6236.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.274 F22Y/A272G isoenzyme A, reduced kcat for NADH Corynebacterium sp.
1.1.1.274 F22Y/K232G/R235G/R238E/A272G isoenzyme A, increase in kcat for NADH Corynebacterium sp.
1.1.1.274 F22Y/K232G/R235G/R238H/A272G isoenzyme A, increase in kcat for NADH Corynebacterium sp.
1.1.1.274 F22Y/K232G/R235T/R238E/A272G isoenzyme A, 24fold increase in kcat for NADH Corynebacterium sp.
1.1.1.274 F22Y/K232G/R238H/A272G isoenzyme A, increase in kcat forNADH Corynebacterium sp.
1.1.1.274 F22Y/S232T/R235S/R238H/A272G isoenzyme A, increase in kcat for NADH Corynebacterium sp.
1.1.1.274 K232G/R238H isoenzyme A, increase in kcat for NADH Corynebacterium sp.
1.1.1.346 F22Y/A272G substrate-binding pocket double mutant with decreased kcat value for NADPH compared to the wild type enzyme Corynebacterium sp.
1.1.1.346 F22Y/K232G/R235G/R238H/A272G mutant with wild type kcat value for NADPH Corynebacterium sp.
1.1.1.346 F22Y/K232G/R235T/R238H/A272G 420 mutant with decreased kcat value for NADPH compared to the wild type enzyme Corynebacterium sp.
1.1.1.346 F22Y/K232G/R238H/A272G mutant with decreased kcat value for NADPH compared to the wild type enzyme Corynebacterium sp.
1.1.1.346 F22Y/K232G/R238H/A272G the mutant exhibits activity with NADH that is more than 2 orders of magnitude higher than that of the wild type enzyme and retains a high level of activity with NADPH Corynebacterium sp.
1.1.1.346 K232G/R238H mutant with decreased kcat value for NADPH compared to the wild type enzyme Corynebacterium sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.274 0.039
-
 NADPH F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.055
-
 NADPH F22Y/S233T/R235S/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.058
-
 NADPH F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.071
-
 NADPH F22Y/K232G/R235T/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.13
-
 NADPH K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.15
-
 NADPH F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.39
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235T/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 0.66
-
 NADH K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 1.2
-
 NADH F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 1.4
-
 NADH isoenzyme A, wild-type Corynebacterium sp.
1.1.1.274 2.4
-
 NADH F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 2.7
-
 NADH F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 3.4
-
 NADH F22Y/K232G/R235G/R238E/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 3.9
-
 NADH F22Y/S233T/R235S/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 5.2
-
 NADH F22Y/K232G/R235T/R238E/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 7.2
-
 2,5-didehydro-D-gluconate F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 8.7
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235T/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 9.1
-
 2,5-didehydro-D-gluconate F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 13
-
 2,5-didehydro-D-gluconate K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 13
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235G/R238E/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 27
-
 2,5-didehydro-D-gluconate F22Y/S232T/R235S/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 30
-
 2,5-didehydro-D-gluconate F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 32
-
 2,5-didehydro-D-gluconate F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 32
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 43
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 110
-
 2,5-didehydro-D-gluconate K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 130
-
 2,5-didehydro-D-gluconate isoenzyme A, wild-type Corynebacterium sp.
1.1.1.274 150
-
 2,5-didehydro-D-gluconate F22Y/K232G/R235T/R238E/A272G isoenzyme A mutant Corynebacterium sp.

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.274 Corynebacterium sp.
-
-
-
1.1.1.346 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
-
 Corynebacterium sp. 2-keto-L-gulonate + NAD+
-
 ?
1.1.1.346 2,5-didehydro-D-gluconate + NADPH + H+
-
 Corynebacterium sp. 2-dehydro-L-gulonate + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.346 2,5-diketo-D-gluconic acid 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 1.23
-
 NADH F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 2.17
-
 NADPH F22Y/S233T/R235S/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 2.83
-
 NADH isoenzyme A, wild-type Corynebacterium sp.
1.1.1.274 3.33
-
 NADPH K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 7
-
 NADPH F22Y/K232G/R235T/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 7.33
-
 NADPH F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 8.5
-
 NADH F22Y/S233T/R235S/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 10
-
 NADH F22Y/K232G/R235G/R238E/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 16.5
-
 NADH F22Y/K232G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 18.3
-
 NADPH F22Y/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 18.3
-
 NADPH F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 18.3
-
 NADH F22Y/K232G/R235G/R238H/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.274 20
-
 NADH K232G/R238H isoenzyme A mutant Corynebacterium sp.
1.1.1.274 68.3
-
 NADH F22Y/K232G/R235T/R238E/A272G isoenzyme A mutant Corynebacterium sp.
1.1.1.346 2.2
-
 NADPH mutant enzyme F22Y/S233T/R235S/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.
1.1.1.346 3.3
-
 NADPH mutant enzyme F22Y/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.
1.1.1.346 7
-
 NADPH mutant enzyme F22Y/K232G/R235G/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.
1.1.1.346 7.3
-
 NADPH mutant enzyme K232G/R238H, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.
1.1.1.346 18.3
-
 NADPH mutant enzyme F22Y/K232G/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.
1.1.1.346 18.3
-
 NADPH wild type enzyme, in 100 mM Bis-Tris, pH 7.0, at 25°C Corynebacterium sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.346 NADPH the enzyme has a preference for NADPH over NADH Corynebacterium sp.