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Information on EC 1.1.1.346 - 2,5-didehydrogluconate reductase (2-dehydro-L-gulonate-forming)
for references in articles please use BRENDA:EC1.1.1.346
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EC Tree 1 Oxidoreductases
1.1 Acting on the CH-OH group of donors
1.1.1 With NAD+ or NADP+ as acceptor
1.1.1.346 2,5-didehydrogluconate reductase (2-dehydro-L-gulonate-forming)
IUBMB Comments
The enzyme is involved in ketogluconate metabolism, and catalyses the reaction in vivo in the reverse direction to that shown . It is used in the commercial microbial production of ascorbate. cf. EC 1.1.1.274, 2,5-didehydrogluconate reductase (2-dehydro-D-gluconate-forming).
The enzyme appears in viruses and cellular organisms
showSynonyms
25dkgr-a, 2,5-dkgr a, 2,5-diketo-d-gluconic acid reductase a, more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
2,5-diketo-D-gluconate reductase
2,5-diketo-D-gluconate reductase I
2,5-diketo-D-gluconate reductase II
2,5-diketo-D-gluconic acid reductase
2,5-diketo-D-gluconic acid reductase A
2,5-DKG reductase
2,5-DKGR A
2,5DKGR
25DKG reductase
2KR
Dkr
YqhE
25DKG reductase
Brevibacterium ketosoreductum ATCC 21914
-
-
-
2KR
Brevibacterium ketosoreductum ATCC 21914
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2-dehydro-L-gulonate + NADP+ = 2,5-didehydro-D-gluconate + NADPH + H+
-
-
-
-
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PATHWAY SOURCE
PATHWAYS
BRENDA
MetaCyc
ketogluconate metabolism, L-ascorbate biosynthesis VIII (engineered pathway)
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SYSTEMATIC NAME
IUBMB Comments
2-dehydro-D-gluconate:NADP+ 2-oxidoreductase (2-dehydro-L-gulonate-forming)
The enzyme is involved in ketogluconate metabolism, and catalyses the reaction in vivo in the reverse direction to that shown [1]. It is used in the commercial microbial production of ascorbate. cf. EC 1.1.1.274, 2,5-didehydrogluconate reductase (2-dehydro-D-gluconate-forming).
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-dehydro-D-gluconate + NADPH + H+
D-gluconate + NADP+
Brevibacterium ketosoreductum
-
Substrates: 11.8% activity compared to 2,5-didehydro-D-gluconate
Products: -
Products: -
?
2-dehydro-L-gulonate + NADPH + H+
L-idonate + NADP+
Brevibacterium ketosoreductum
-
Substrates: 10.8% activity compared to 2,5-didehydro-D-gluconate
Products: -
Products: -
?
5-dehydro-D-fructose + NADPH + H+
? + NADP+
-
Substrates: -
Products: -
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?
ethyl 2-acetylpent-4-enoate + NADPH + H+
ethyl (2R)-2-[(1S)-1-hydroxyethyl]pent-4-enoate + NADP+
-
Substrates: 250% activity compared to ethyl 2-methylacetoacetate
Products: -
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?
ethyl 2-ethyl-3-oxobutanoate + NADPH + H+
ethyl (2R,3S)-2-ethyl-3-hydroxybutanoate + NADP+
-
Substrates: 120% activity compared to ethyl 2-methylacetoacetate
Products: -
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?
ethyl 2-methylacetoacetate + NADH + H+
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NAD+
-
Substrates: 7% activity with NADH compared to NADPH
Products: -
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?
ethyl 2-methylacetoacetate + NADPH + H+
ethyl (2R)-methyl-(3S)-hydroxybutanoate + NADP+
-
Substrates: -
Products: -
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?
ethyl acetoacetate + NADPH + H+
ethyl (3S)-3-hydroxybutanoate + NADP+
-
Substrates: 53% activity compared to ethyl 2-methylacetoacetate
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
Substrates: -
Products: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: isoform DKGR B exhibits 66fold higher specific activity toward 2,5-didehydro-D-gluconate than isoform DKGR A
Products: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: a reduction of substrate by NADPH is highly preferred
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r
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: 100% activity
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: a reduction of substrate by NADPH is highly preferred
Products: -
Products: -
r
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
-
Substrates: 100% activity
Products: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
2-dehydro-L-gulonate + NADP+
Substrates: NADPH is the preferred electron donor
Products: -
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?
2,5-didehydro-D-gluconate + NADPH + H+
5-dehydro-D-gluconate + NADP+
Brevibacterium ketosoreductum
-
Substrates: 100% activity
Products: -
Products: -
?
2,5-didehydro-D-gluconate + NADPH + H+
5-dehydro-D-gluconate + NADP+
Brevibacterium ketosoreductum ATCC 21914
-
Substrates: 100% activity
Products: -
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?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
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?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
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?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
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?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
r
5-dehydro-D-fructose + NADPH + H+
L-sorbose + NADP+
-
Substrates: isoforms 2,5-diketo-D-gluconate reductase I and II show 150% and 13% activity, respectively, compared to 2,5-didehydro-D-gluconate
Products: -
Products: -
?
5-dehydro-D-fructose + NADPH + H+
L-sorbose + NADP+
-
Substrates: isoforms 2,5-diketo-D-gluconate reductase I and II show 150% and 13% activity, respectively, compared to 2,5-didehydro-D-gluconate
Products: -
Products: -
?
additional information
?
-
Brevibacterium ketosoreductum
-
Substrates: no activity with D-gluconate, D-fructose, L-sorbose, and 5-dehydro-D-gluconate
Products: -
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?
additional information
?
-
-
Substrates: the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
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?
additional information
?
-
-
Substrates: the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: no activity with 2-dehydro-L-gulonate, 2-dehydro-D-gluconate, 5-dehydro-D-gluconate, D-fructose, and L-sorbose
Products: -
Products: -
?
additional information
?
-
-
Substrates: the wild type enzyme shows no activity with NADH
Products: -
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?
additional information
?
-
-
Substrates: 2-keto-D-gluconate, 2-keto-L-gulonate, 5-keto-D-gluconate, D-fructose, and L-sorbose are inactive when assayed at pH 6.0 in the presence of NADH or NADPH
Products: -
Products: -
?
additional information
?
-
Substrates: 2-keto-D-gluconate, 2-keto-L-gulonate, 5-keto-D-gluconate, D-fructose, and L-sorbose are inactive when assayed at pH 6.0 in the presence of NADH or NADPH
Products: -
Products: -
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
LITERATURE
COMMENTARY
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
?
2-dehydro-L-gulonate + NADP+
2,5-didehydro-D-gluconate + NADPH + H+
-
Substrates: -
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
Products: -
Products: -
?
additional information
?
-
-
Substrates: the enzyme catalyzes degradation of estradiol, oestrone, testosterone, and methyltestosterone, overview
Products: -
Products: -
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.8
2,5-didehydro-D-gluconate
-
isoform 2,5-diketo-D-gluconate reductase I, at pH 7.0 and 30°C
13.5
2,5-didehydro-D-gluconate
-
isoform 2,5-diketo-D-gluconate reductase II, at pH 7.0 and 30°C
0.012
NADPH
-
isoform 2,5-diketo-D-gluconate reductase I, at pH 7.0 and 30°C
0.013
NADPH
-
isoform 2,5-diketo-D-gluconate reductase II, at pH 7.0 and 30°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.2
NADPH
-
mutant enzyme F22Y/S233T/R235S/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C
3.3
NADPH
-
mutant enzyme F22Y/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C
7
NADPH
-
mutant enzyme F22Y/K232G/R235G/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C
7.3
NADPH
-
mutant enzyme K232G/R238H, in 100 mM Bis-Tris, pH 7.0, at 25°C
18.3
NADPH
-
mutant enzyme F22Y/K232G/R238H/A272G, in 100 mM Bis-Tris, pH 7.0, at 25°C
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.188
-
recombinant enzyme, expressed in Lactococcus lactis strain NZ3900 from plasmid pVK51dkr induced in pH-regulated cultivation, pH 6.5, 22°C
0.191
-
recombinant enzyme, expressed in Lactococcus lactis strain NZ3900 from plasmid pVK51dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C
0.232
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP603dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C
0.243
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP609dkr induced in not-pH-regulated cultivation, pH 6.5, 22°C
0.264
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP603dkr induced in pH-regulated cultivation, pH 6.5, 22°C
0.308
-
recombinant enzyme, expressed in Lactobacillus plantarum strain TLG02 from plasmid pSIP609dkr induced in pH-regulated cultivation, pH 6.5, 22°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.7
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Highest Expressing Human Cell Lines
Cell Line LinksPlease wait a moment until the data is sorted. This message will disappear when the data is sorted.
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
-
compared to the wild-type, the knockout mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone. Cell growth on ethanol, oestrone, estradiol, testosterone or methyltestosterone is reduced in the mutant strain compared to the wild-type
metabolism
physiological function
additional information
evolution
-
the enzyme belongs to the AKR superfamily, monomeric (alpha/beta) 8-barrel proteins which bind NAD(P)(H) to metabolize an array of substrates
evolution
-
the enzyme belongs to the AKR superfamily, monomeric (alpha/beta) 8-barrel proteins which bind NAD(P)(H) to metabolize an array of substrates
-
metabolism
-
the enzyme reduces 2,5-didehydro-D-gluconate, a key step in the microbial synthesis of vitamin C
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
metabolism
-
the enzyme reduces 2,5-didehydro-D-gluconate, a key step in the microbial synthesis of vitamin C
-
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
-
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
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
-
additional information
-
three consensus sequences of the AKR superfamily are found as GxxxxDxAxxY, LxxxGxxxPxxGxG and LxxxxxxxxxDxxxxH. GxxxxDxAxxY is the active site, LxxxGxxxPxxGxG is the cofactor-binding site for NAD(P)(H), and LxxxxxxxxxDxxxxH is required for supporting the 3D structure
additional information
-
three consensus sequences of the AKR superfamily are found as GxxxxDxAxxY, LxxxGxxxPxxGxG and LxxxxxxxxxDxxxxH. GxxxxDxAxxY is the active site, LxxxGxxxPxxGxG is the cofactor-binding site for NAD(P)(H), and LxxxxxxxxxDxxxxH is required for supporting the 3D structure
-
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). DKGA_CORSC
278
0
30119
Swiss-Prot
-
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
29000
34000
35000
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
monomer
?
-
x * 39400, about, sequence calculation, x * 41600, recombinant His-tagged enzyme, SDS-PAGE
?
-
x * 39400, about, sequence calculation, x * 41600, recombinant His-tagged enzyme, SDS-PAGE
-
homodimer
Brevibacterium ketosoreductum ATCC 21914
-
2 * 35000, SDS-PAGE
-
monomer
-
1 * 29000, isoform 2,5-diketo-D-gluconate reductase I, SDS-PAGE
monomer
-
1 * 34000, isoform 2,5-diketo-D-gluconate reductase II, SDS-PAGE
monomer
-
1 * 29000, isoform 2,5-diketo-D-gluconate reductase I, SDS-PAGE
-
monomer
-
1 * 34000, isoform 2,5-diketo-D-gluconate reductase II, SDS-PAGE
-
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with NADPH, hanging drop vapor diffusion method, using 1 M sodium phosphate, 1 M potassium phosphate, 100 mM HEPES buffer, pH 7.0, at 4°C
-
mutant enzyme F22Y/K232G/R238H/A272G in complex with NADH, hanging drop vapor diffusion method, using 1.5 M lithium sulfate and 0.1 M Na HEPES (pH 7.5), at 22°C
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
F22Y
-
the mutation causes a 2.5fold decrease in Km for 2,5-didehydro-D-gluconate whereas the value of kcat remains essentially unchanged
F22Y/A272G
-
substrate-binding pocket double mutant with decreased kcat value for NADPH compared to the wild type enzyme
F22Y/K232G/R235G/R238H/A272G
-
mutant with wild type kcat value for NADPH
F22Y/K232G/R235T/R238H/A272G 420
-
mutant with decreased kcat value for NADPH compared to the wild type enzyme
F22Y/K232G/R238H/A272G
K232G/R238H
-
mutant with decreased kcat value for NADPH compared to the wild type enzyme
K233G
-
the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme
K233H
-
the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity
K233M
-
the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme
K233Q
-
the mutant shows wild type NADPH activity and increased NADH activity
K233R
-
the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity
K233S
-
the mutant shows wild type NADPH activity and increased NADH activity compared to the wild type enzyme
Q192R
-
the mutation primarily affects the kcat parameter toward the 2,5-didehydro-D-gluconate substrate, increasing its value approximately 2.5fold, whereas Km is relatively unaffected, or increases slightly
R235D
-
the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity
R235E
-
the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity
R235G
-
the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme
R235T
-
the mutant shows wild type NADPH activity and increased NADH activity
R235Y
-
the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity
R238E
-
the mutant shows no activity with NADPH and increased NADH activity compared to the wild type enzyme
R238G
-
the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity
R238H
-
the mutant shows wild type NADPH activity and increased NADH activity
R238Q
-
the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity
R238Y
-
the mutant shows reduced NADPH activity and increased NADH activity ompared to the wild type enzyme
V234M/R235C
-
the mutant shows wild type NADPH activity and no NADH activity
V234S
-
the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity
additional information
F22Y/K232G/R238H/A272G
the mutation enhances binding to NADH, while retaining to a large extent the ability to bind NADPH. The mutant is also more stable and can, therefore, be expected to exhibit greater effective activity at elevated temperatures in comparison to the wild type enzyme
F22Y/K232G/R238H/A272G
-
mutant with decreased kcat value for NADPH compared to the wild type enzyme
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
additional information
-
construction of enzyme gene knockout mutant M-AKR, that shows decreased degradation activity with testosterone, estradiol, oestrone, and methyltestosterone compared to the wild-type enzyme. Compared to the wild-type, the mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone
additional information
-
construction of enzyme gene knockout mutant M-AKR, that shows decreased degradation activity with testosterone, estradiol, oestrone, and methyltestosterone compared to the wild-type enzyme. Compared to the wild-type, the mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone
-
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
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
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 7
-
when isoform I is incubated at different pH values for 3 h at 30°C, the remaining activity is about 29% of the original level at pH 5.0, about 65% at pH 6.0 and about 51% at pH 7.0
5.2 - 10
Brevibacterium ketosoreductum
-
after 2 h incubation at 30°C, the remaining activity is about 35% of the original level at pH 8.4, and no remaining activity is observed below pH 5.1 and 10.0. The enzyme is stable from pH 6.0 to 7.5
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0 - 38
-
temperature stability of isoform 2,5-diketo-D-gluconate reductase I is measured by storing the enzyme in 0.1 M Tris-HC1 buffer (pH 7.0) for 1 h. The activity of the enzyme decreases gradually with temperature from 0°C to 27°C, falls rapidly above 27°C, and is lost completely at about 38°C. The remaining activity at 27°C is about 77% of that at 0°C
40 - 80
-
after approximately 10 min of deactivation, enzyme activity attains a constant value, independently of deactivation temperature (40-80°C). This value corresponds to the 30% of native activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, 2 mg/ml enzyme in 20 mM Tris-HC1 (pH 7.5), 6 months, the enzyme remains stable
-
4°C, enzyme solution at pH 6.5-7.5, at least 2 months, the enzyme remains stable
-
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE Toyopearl column chromatography and phenyl Toyopearl column chromatography, Blue Sepharose column chromatography and Sephacryl S-200 gel filtration
Brevibacterium ketosoreductum
-
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and Amicon Matrix Gel Red A gel filtration
-
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, Matrex Red-A column chromatography, Sephacryl S-200 gel filtration, and hypatite C cholumn chromatography
-
DEAE-cellulose column chromatography, Cibacron blueF 3GA affinity column chromatography, and and TSK gel filtration
-
DEAE-Sephacel column chromatography and Cibacron blue 3GA column chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene CTATCC11996_22452, genetic organization, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
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
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme expression is significantly induced by estrone, estradiol, and methyltestosterone, and slightly by progesterone, but not by testosterone
the enzyme expression is significantly induced by estrone, estradiol, and methyltestosterone, and slightly by progesterone, but not by testosterone
-
the enzyme expression is significantly induced by estrone, estradiol, and methyltestosterone, and slightly by progesterone, but not by testosterone
-
-
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
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
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
-
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Yum, D-Y.; Bae, S-S.; Pan, J-G.
Purification and characterization of the 2-ketoaldonate reductase from Brevibacterium ketosoreductum ATCC21914
Biosci. Biotechnol. Biochem.
62
154-156
1998
Brevibacterium ketosoreductum, Brevibacterium ketosoreductum ATCC 21914
brenda
Sonoyama, T.; Kobayashi, K.
Purification and properties of two 2,5-diketo-D-gluconate reductases from a mutant strain derived from Corynebacterium sp
J. Ferment. Technol.
65
311-317
1987
Corynebacterium sp., Corynebacterium sp. SHS 0007
-
brenda
Miller, J.V.; Estell, D.A.; Lazarus, R.A.
Purification and characterization of 2,5-diketo-D-gluconate reductase from Corynebacterium sp
J. Biol. Chem.
262
9016-9020
1987
Corynebacterium sp., Corynebacterium sp. ATCC 31090
brenda
Khurana, S.; Powers, D.B.; Anderson, S.; Blaber, M.
Crystal structure of 2,5-diketo-D-gluconic acid reductase A complexed with NADPH at 2.1 A resolution
Proc. Natl. Acad. Sci. USA
95
6768-6773
1998
Corynebacterium sp.
brenda
Yum, D.Y.; Lee, B.Y.; Pan, J.G.
Identification of the yqhE and yafB genes encoding two 2,5-diketo-D-gluconate reductases in Escherichia coli
Appl. Environ. Microbiol.
65
3341-3346
1999
Escherichia coli, Escherichia coli (Q46857)
brenda
Khurana, S.; Sanli, G.; Powers, D.B.; Anderson, S.; Blaber, M.
Molecular modeling of substrate binding in wild-type and mutant Corynebacteria 2,5-diketo-D-gluconate reductases
Proteins
39
68-75
2000
Corynebacterium sp.
brenda
Banta, S.; Swanson, B.A.; Wu, S.; Jarnagin, A.; Anderson, S.
Alteration of the specificity of the cofactor-binding pocket of Corynebacterium 2,5-diketo-D-gluconic acid reductase A
Protein Eng.
15
131-140
2002
Corynebacterium sp.
brenda
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
Biochemistry
41
6226-6236
2002
Corynebacterium sp.
brenda
Habrych, M.; Rodriguez, S.; Stewart, J.D.
Purification and identification of an Escherichia coli beta-keto ester reductase as 2,5-diketo-D-gluconate reductase YqhE
Biotechnol. Prog.
18
257-261
2002
Escherichia coli, Escherichia coli BL21-(DE3)
brenda
Sanli, G.; Banta, S.; Anderson, S.; Blaber, M.
Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase
Protein Sci.
13
504-512
2004
Corynebacterium sp. (P06632)
brenda
Maremonti, M.; Greco Jr., G.; Wichmann, R.
Characterisation of 2,5-diketo-D-gluconic acid reductase from Corynebacterium sp.
Biotechnol. Lett.
18
845-850
1996
Corynebacterium sp.
-
brenda
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
AMB Express
3
7-7
2013
Corynebacterium glutamicum, Corynebacterium glutamicum DSM 20301
brenda
Chen, Y.; Ji, W.; Zhang, H.; Zhang, X.; Yu, Y.
Cloning, expression and characterization of a putative 2,5-diketo-D-gluconic acid reductase in Comamonas testosteroni
Chem. Biol. Interact.
234
229-235
2015
Comamonas testosteroni, Comamonas testosteroni ATCC 11996
brenda
EXTERNAL LINKS (specific for EC-Number 1.1.1.346)
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