Information on EC 1.1.1.69 - gluconate 5-dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.1.1.69
-
RECOMMENDED NAME
GeneOntology No.
gluconate 5-dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-gluconate + NAD(P)+ = 5-dehydro-D-gluconate + NAD(P)H + H+
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
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redox reaction
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-
-
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reduction
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
L-idonate degradation
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ketogluconate metabolism
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SYSTEMATIC NAME
IUBMB Comments
D-gluconate:NAD(P)+ 5-oxidoreductase
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CAS REGISTRY NUMBER
COMMENTARY hide
9028-70-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
IFO 3281 and IFO 3284
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-
Manually annotated by BRENDA team
Acetobacter acetigenus
IFO 3277
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-
Manually annotated by BRENDA team
Acetobacter ascendens
IFO 3299
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Manually annotated by BRENDA team
IFO 3129
-
-
Manually annotated by BRENDA team
Acetobacter kuetzingianus, IFO 3222; IFO 3298
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-
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
IFO 3245 and IFO 3247
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-
Manually annotated by BRENDA team
IFO 3250
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-
Manually annotated by BRENDA team
IFO 3262, IFO 3265 and IFO 3267
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-
Manually annotated by BRENDA team
Gluconobacter gluconicus
IFO 3171
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Manually annotated by BRENDA team
strain DSM2343 (ATCC 621H)
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Manually annotated by BRENDA team
IFO 3990
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-
Manually annotated by BRENDA team
IFO 12467
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Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
IFO 3288
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,3-butanediol + NAD+
? + NADH
show the reaction diagram
5-dehydro-D-fructose + NADPH
D-fructose + NADP+
show the reaction diagram
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23% of the activity with 5-dehydro-D-gluconate
-
-
?
5-dehydro-D-gluconate + NADH
D-gluconate + NAD+
show the reaction diagram
5-dehydro-D-gluconate + NADH + H+
D-gluconate + NAD+
show the reaction diagram
5-dehydro-D-gluconate + NADPH
D-gluconate + NADP+
show the reaction diagram
5-dehydro-D-gluconate + NADPH + H+
D-gluconate + NADP+
show the reaction diagram
5-keto D-gluconate + NADPH
D-gluconate + NADP+
show the reaction diagram
-
-
-
-
?
D-arabitol + NAD+
? + NADH
show the reaction diagram
D-fructose + NADPH
5-dehydro-D-fructose + NADP+
show the reaction diagram
D-gluconate + NAD+
5-dehydro-D-gluconate + NADH + H+
show the reaction diagram
D-gluconate + NADP+
5-dehydro-D-gluconate + NADPH
show the reaction diagram
D-gluconate + NADP+
5-dehydro-D-gluconate + NADPH + H+
show the reaction diagram
D-gluconic acid + NAD+
? + NADH
show the reaction diagram
D-glucono-delta-lactone + NADP+
?
show the reaction diagram
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14% of the activity with D-gluconate
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-
?
D-glucose + ?
5-keto-D-gluconic acid + ?
show the reaction diagram
D-mannitol + NAD+
? + NADH
show the reaction diagram
D-sorbitol + NAD+
? + NADH
show the reaction diagram
glycerol + NAD+
? + NADH
show the reaction diagram
138% of activity with D-sorbitol
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-
?
meso-erytritol + NAD+
? + NADH
show the reaction diagram
143% of activity with D-sorbitol
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?
ribitol + NAD+
? + NADH
show the reaction diagram
66% of activity with D-sorbitol
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-
?
sorbitol + NAD+
? + NADH
show the reaction diagram
additional information
?
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production of 5-keto D-gluconate from D-gluconate
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
5-dehydro-D-gluconate + NADH + H+
D-gluconate + NAD+
show the reaction diagram
5-dehydro-D-gluconate + NADPH
D-gluconate + NADP+
show the reaction diagram
5-dehydro-D-gluconate + NADPH + H+
D-gluconate + NADP+
show the reaction diagram
D-gluconate + NAD+
5-dehydro-D-gluconate + NADH + H+
show the reaction diagram
D-gluconate + NADP+
5-dehydro-D-gluconate + NADPH + H+
show the reaction diagram
additional information
?
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production of 5-keto D-gluconate from D-gluconate
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
1 mM, slight activation
Fe3+
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1 mM, activation
Li+
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1 mM, slight activation
Mg2+
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1 mM, slight activation
Mn2+
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1 mM, increases the activity to 127%
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cd2+
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1 mM, 29% inhibition
Cu2+
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1 mM, 40% loss of activity
NEM
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1 mM, 82% inhibition
Sn2+
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1 mM, 20% inhibition
sodium lauryl sulfate
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1 mM, 76% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyrroloquinoline quinone
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.9
5-Dehydro-D-gluconate
4.6
D-arabitol
pH 5.5, 30°C
18 - 20.6
D-gluconate
161
D-gluconic acid
pH 5.5, 30°C
57
D-sorbitol
pH 5.5, 30°C
0.016 - 0.073
NADP+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
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reduction of 5-dehydro-D-gluconate
5.5
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reduction of 5-dehydro-D-gluconate
6 - 7
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reduction of 5-dehydro-D-gluconate
6 - 7
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reduction of 5-dehydro-D-gluconate
7.3 - 7.5
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reduction of 5-dehydro-D-gluconate
7.5
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reduction of 5-dehydro-D-gluconate
9.3
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oxidation of D-gluconate
10 - 11
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oxidation of D-gluconate
10 - 11
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oxidation of D-gluconate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 8.5
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pH 4.5: about 70% of maximal activity, pH 8.5: about 60% of maximal activity, reduction of 5-dehydro-D-gluconate
8 - 12
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pH 8.0: about 30% of maximal activity, pH 12.0: 35% of maximal activity, oxidation of D-gluconate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Burkholderia cenocepacia (strain ATCC BAA-245 / DSM 16553 / LMG 16656 / NCTC 13227 / J2315 / CF5610)
Streptococcus suis (strain 05ZYH33)
Streptococcus suis (strain 05ZYH33)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000
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4 * 25000, SDS-PAGE
29000
4 * 29000, X-ray diffraction and SDS-PAGE
75000
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gel filtration
80000
x * 80000, SDS-PAGE
100000
110000
132000
native-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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2 * 75000, SDS-PAGE
homotetramer
4 * 29000, X-ray diffraction and SDS-PAGE
tetramer
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4 * 25000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
fairly stable
654316
8
unstable
654316
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
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loss of activity after a few min
50
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pH 7.5, 10 min, stable up to
55
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5 min, in presence of D-gluconate or 5-dehydro-D-gluconate, even at 0.0001 mg/ml protein
70
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10 min, pH 7.5, about 75% loss of activity
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
75% loss of activity within a few days unless either D-gluconate or 5-dehydro-D-gluconate is added during storage or purification
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unstable to dialysis in a cold room, in presence of sulfhydryl compounds, EDTA, sucrose and/or glycerol
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 10 mM phosphate buffer, pH 6.0, no added 2-mercaptoethanol, fully stable for at least 3 days
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5°C, pH 7.5, 0.01 M potassium phosphate buffer, 3 d, complete loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
glutathione Sepharose column chromatography and Superdex 200 gel filtration
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recombinant protein
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as fusion protein with a N-terminal His6 tag in Escherichia coli strain BL21 (DE3)
expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21(DE3), expression levels vary from colony to colony, cells showing high enzyme expression tend to show less growth, expression system appears to be unstable as expression level decreases when subcultured for several times, best expression results with freshly transformed cells
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expression in Escherichia coli and Gluconobacter oxydans
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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enzymatic quantification of 5-keto-D-gluconate
food industry
synthesis
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strain overexpressing enzyme plus Escherichia coli transhydrogenase sthA, enhanced accumulation of 5-ketoD-gluconate, precursor of L-(+)-tartaric acid
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