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Literature summary for 1.1.1.346 extracted from

  • Kaswurm, V.; Nguyen, T.T.; Maischberger, T.; Kulbe, K.D.; Michlmayr, H.
    Evaluation of the food grade expression systems NICE and pSIP for the production of 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum (2013), AMB Express, 3, 7-17 .
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis 2,5-DKG reductase is an integral part of several industrial processes designed to synthesize 2-dehydro-L-gulonate based on the 2,5-diketo-D-gluconic acid pathway Corynebacterium glutamicum

Cloned(Commentary)

Cloned (Comment) Organism
gene dkr, recombinant expression in Lactococcus lactis strain NZ3900, and Lactobacillus plantarum strains WCFS1 and TLG02, cloning in Escherichia coli strains MB2159 and NEB 5-alpha, expression method evaluation Corynebacterium glutamicum

Engineering

Protein Variants Comment Organism
additional information establishment of an efficient process for 2,5-DKG reductase production that also satisfies food safety requirements. Food grade variants of the Lactobacillales based expression systems pSIP (Lactobacillus plantarum) and NICE (Lactococcus lactis) are evaluated with regard to their effictiveness to produce 2,5-DKG reductase from Corynebacterium glutamicum, overview. Lactobacillus plantarum/pSIP609 is an interesting alternative to Escherichia coli expression systems for industrial 2,5-DKG reductase production. Highest production levels of 2,5-DKG reductase are obtained with the system Lactobacillus plantarum/pSIP609, resulting in 104 U/l without pH regulation and 262 U/l with pH control at pH 6.5 Corynebacterium glutamicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-dehydro-L-gulonate + NADP+ Corynebacterium glutamicum
-
2,5-didehydro-D-gluconate + NADPH + H+
-
?
2-dehydro-L-gulonate + NADP+ Corynebacterium glutamicum DSM 20301
-
2,5-didehydro-D-gluconate + NADPH + H+
-
?

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
-
-
Corynebacterium glutamicum DSM 20301
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.188
-
recombinant enzyme, expressed in Lactococcus lactis strain NZ3900 from plasmid pVK51dkr induced in pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum
0.191
-
recombinant enzyme, expressed in Lactococcus lactis strain NZ3900 from plasmid pVK51dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum
0.232
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP603dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum
0.243
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP609dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum
0.264
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP603dkr induced in pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum
0.308
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP609dkr induced in pH-regulated cultivation, pH 6.5, 22°C Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-dehydro-L-gulonate + NADP+
-
Corynebacterium glutamicum 2,5-didehydro-D-gluconate + NADPH + H+
-
?
2-dehydro-L-gulonate + NADP+
-
Corynebacterium glutamicum DSM 20301 2,5-didehydro-D-gluconate + NADPH + H+
-
?

Subunits

Subunits Comment Organism
? x * 31000, recombinant enzyme, SDS-PAGE Corynebacterium glutamicum

Synonyms

Synonyms Comment Organism
2,5-diketo-D-gluconic acid reductase
-
Corynebacterium glutamicum
2,5-DKG reductase
-
Corynebacterium glutamicum
Dkr
-
Corynebacterium glutamicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at room temperature Corynebacterium glutamicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
assay at Corynebacterium glutamicum

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Corynebacterium glutamicum
NADPH
-
Corynebacterium glutamicum

General Information

General Information Comment Organism
metabolism 2,5-DKG reductase catalyses the stereospecific reduction of 2,5-diketo-D-gluconic acid (2,5-DKG) at position C-5 to 2-keto-L-gulonic acid (2-KLG), a key intermediate in the production of L-ascorbic acid Corynebacterium glutamicum
physiological function the enzyme catalyses the reduction of 2,5-diketo-D-gluconic acid to 2-keto-L-gulonic acid, a direct precursor (lactone) of L-ascorbic acid (vitamin C). This reaction is an essential step in the biocatalytic production of the food supplement vitamin C from D-glucose or D-gluconic acid Corynebacterium glutamicum