Information on EC 1.1.1.119 - glucose 1-dehydrogenase (NADP+)

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

EC NUMBER
COMMENTARY hide
1.1.1.119
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RECOMMENDED NAME
GeneOntology No.
glucose 1-dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-glucose + NADP+ = D-glucono-1,5-lactone + NADPH + H+
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
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redox reaction
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reduction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Entner-Doudoroff pathway II (non-phosphorylative)
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Entner-Doudoroff pathway III (semi-phosphorylative)
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Entner Doudoroff pathway
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SYSTEMATIC NAME
IUBMB Comments
D-glucose:NADP+ 1-oxidoreductase
Also oxidizes D-mannose, 2-deoxy-D-glucose and 2-amino-2-deoxy-D-mannose.
CAS REGISTRY NUMBER
COMMENTARY hide
37250-50-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
recombinant enzyme, enzyme-based synthesis system for poly(3-hydroxybutyrate)
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Manually annotated by BRENDA team
IFO 12528
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Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-amino-2-deoxy-D-mannose + NADP+
2-amino-2-deoxy-D-mannono-1,5-lactone + NADPH
show the reaction diagram
2-deoxy-D-glucose + NADP+
2-deoxy-D-glucono-1,5-lactone + NADPH + H+
show the reaction diagram
beta-D-galactose + NADP+
D-galactono-1,5-lactone + NADPH
show the reaction diagram
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-
-
-
?
beta-D-glucose + NAD+
?
show the reaction diagram
beta-D-glucose + NAD+
D-glucono-1,5-lactone + NADH + H+
show the reaction diagram
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the enzyme has a strong preference for NADP+ over NAD+
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?
beta-D-glucose + NADP+
?
show the reaction diagram
beta-D-glucose + NADP+
D-glucono-1,5-lactone + NADPH + H+
show the reaction diagram
D-glucose + 1,N6-ethanoadenine-NADP+
D-glucono-1,5-lactone + 1,N6-ethanoadenine-NADPH
show the reaction diagram
D-glucose + N1-(2-aminoethyl)-NADP+
D-glucono-1,5-lactone + N1-(2-aminoethyl)-NADPH
show the reaction diagram
D-glucose + N6-(2-aminoethyl)-NADP+
D-glucono-1,5-lactone + N6-(2-aminoethyl)-NADPH
show the reaction diagram
D-glucose + NAD(P)+
D-glucono-1,5-lactone + NAD(P)H + H+
show the reaction diagram
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?
D-glucose + NAD+
D-glucono-1,5-lactone + NADH
show the reaction diagram
D-glucose + NAD+
D-glucono-1,5-lactone + NADH + H+
show the reaction diagram
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?
D-glucose + NADP+
D-glucono-1,5-lactone + NADPH
show the reaction diagram
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?
D-glucose + NADP+
D-glucono-1,5-lactone + NADPH + H+
show the reaction diagram
D-mannose + NADP+
D-mannono-1,5-lactone + NADPH
show the reaction diagram
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
D-glucose + NADP+
D-glucono-1,5-lactone + NADPH + H+
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,N6-ethanoadenine-NADP+
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N1-(2-aminoethyl)-NADP+
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N6-(2-aminoethyl)-NADP+
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NADP+
NADPH
polyethylene glycol(MW 20000)-N6-(2-aminoethyl)-NADP+
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
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the activity is dependent on the concentration of NaCl or KCl in the activity buffer, being optimal at 1.3 M
Mg2+
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besides KCl/NaCl, the activity also depends on presence of bivalent cations. Mg2+ is more effective than Mg2+ or Ni2+
Mn2+
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besides KCl/NaCl, the activity also depends on presence of bivalent cations. Mn2+ is less effective than Mg2+ and more effective than Ni2+
NaCl
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the activity is dependent on the concentration of NaCl or KCl in the activity buffer, being optimal at 1.3 M
Ni2+
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besides KCl/NaCl, the activity also depends on presence of bivalent cations. Ni2+ is more effective than Mg2+ or Mn2+
Zinc
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a member of the zinc-containing medium chain polyol dehydrogenase family
Zn2+
dependent on; zinc is an important component of the catalytic centre
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KCl
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at 40C, half-life in 0.5 M KCl is 86 h for wild-type, and 114 h for mutant D172K/D216K/D344K, 95 h for mutant D172K, 117 h for mutant D216K, 114 h for mutant D344K. At 25C, half-life in 0.5 M KCl is 506 h for wild-type, and 613 h for mutant D172K/D216K/D344K, 630 h for mutant D172K, 660 h for mutant D216K, 537 h for mutant D344K
NADPH
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competitive to NADP+
p-chloromercuribenzoate
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sulfhydryl reagents
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NaCl
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triple mutant D172K/D216K/D344K shows its maximum activity in a buffer with 0.50-0.75 M NaCl while the wild type protein has its maximum activity with 1.5 M NaCl
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
1,N6-ethanoadenine-NADP+
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29
2-amino-2-deoxy-D-mannose
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39
2-deoxy-D-glucose
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3.9 - 80
beta-D-glucose
0.0286 - 80
D-glucose
1.8 - 5.6
D-mannose
0.07
N1-(2-aminoethyl)-NADP+
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0.03
N6-(2-aminoethyl)-NADP+
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0.035 - 0.4
NAD(P)+
0.4 - 4
NAD+
0.00588 - 2.6
NADP+
0.038
NADPH
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0.09
polyethylene glycol(MW 20000)-N6-(2-aminoethyl)-NADP+
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00041 - 16.7
D-glucose
0.085 - 16.7
NADP+
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0000008 - 5.25
D-glucose
35
0.000035 - 0.00112
NAD+
7
0.0001 - 0.893
NADP+
10
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.09 - 2
NAD(P)+
1 - 2.1
NAD+
0.09 - 1.3
NADP+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
550
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pH 8.8, 40C
additional information
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25 mU/mg at 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
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in Tris-buffer
8.5 - 9
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in Tris-buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 9
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broad range of optimal activity, in Tris-buffer consistently higher than that in phosphate-buffers
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
19500
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2 * 19500
40000
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4 * 40000, SDS-PAGE
53000
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2 * 53000, SDS-PAGE
89000
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gel filtration
153000
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gel filtration on Sephadex G-200
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
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4 * 40000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1.6 A structure determination of the enzyme using crystals grown under conditions that closely mimic those experienced within the cell of the halophile. The structure is compared with nonhalophilic homologues. Location of bound counterions in the structure. Analysis of protein-solvent interactions to further the understanding of the molecular basis of halophilic adaptation
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; the structures of GlcDH in binary complexes with either NADP+/Zn2+ or NADPH/Zn2+, a ternary complex with NADP+/Zn2+/glucose, and a nonproductive ternary complex with NADP+/Zn2+ /gluconolactone are determined from isomorphous crystals, in the presence of zinc (1 mM) and either 5 mM NADP+, by using 1.4-1.6 M sodium citrate as the precipitant (in 100 mM HEPES, pH 7.0, and 2 M NaCl) or 5 mM NADPH, using 67-72% (w/v) 2-methyl-2,4-pentanediol as precipitant (in 100 mM HEPES, pH 7.5, and 2 M NaCl)
hanging-drop vapour diffusion; hanging-drop vapour-diffusion method at 17C, crystallization of a range of binary and ternary complexes of the wild-type and a D38C mutant protein. For most of the complexes, crystals belonging to space group I222 are obtained using sodium/potassium citrate as a precipitant. For the binary and non-productive ternary complexes with NADPH/Zn, it is necessary to replace the citrate with 2-methyl-2,4-pentanediol. Despite the radical change in conditions, the crystals thus formed are isomorphous
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hanging-drop vapour-diffusion method using sodium citrate as the precipitant. Two crystal forms representing the free enzyme and the binary complex with NADP+ grow under these conditions. Crystals of form I diffract to beyond 3.5 A resolution and belong to the hexagonal space group P622, with unit-cell parameters a = b = 89.1, c = 214.6 A, alpha = beta = 90, gamma = 120. Crystals of form II diffract to greater than 2.0 A and belong to the orthorhombic space group I222 or I212121, with unit-cell parameters a = 61.8, b = 110.9, c = 151.7 A, alpha = beta = gamma = 90
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
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half-life: 56.8 h, in the presence of 3 M NaCl, stability decreases significantly with lower salt concentrations
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
freezing and thawing: significant loss of activity
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lyophilization of dilute enzyme solutions, below 0.5 ng/ml, results in significant loss of activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 1 year
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4C, pH 5.4 - pH 8.5, 8 weeks
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4C, stable for several months
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4C, stable for weeks
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5C, crystalline preparation in ammonium sulfate, 6 months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
column chromatography on DEAE-Sephadex A-50 and affinty chromatography by blue-dextran Sepharose 4B
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as recombinant protein in Escherichia coli strain BL21(DE3); expressed in Escherichia coli BL21 (DE3) cells; expression as inclusion bodies in Escherichia coli
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expression in Escherichia coli
expression in Escherichia coli BL21 (DE3) already heterologously overexpressing P450 BM-3 QM together with glucose facilitator (GLF) from Zymomonas mobilis; expression in Escherichia coli, together with a glucose facilitator from Zymomonas mobilis for uptake of unphosphorylated glucose
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overexpressed in Escherichia coli
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overexpression in Escherichia coli JM109 for use as a cofactor regenerator
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D172K
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mutation in surface residue, mutant protein is slightly less halotolerant than wild-type
D172K/D216K/D344K
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mutation in surface residue, mutant protein is slightly less halotolerant than wild-type
D216K
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mutation in surface residue, mutant protein is slightly less halotolerant than wild-type
D344K
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mutation in surface residue, mutant protein is slightly less halotolerant than wild-type
D38C
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crystallization of a range of binary and ternary complexes of the wild-type and a D38C mutant protein
G206D
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less efficient with NADP+ than the wild type, 1.6fold increase in ratio kcat/Km for NAD+; prefers NAD+ over NADP+; the mutant is less efficient with NADP+ than the wild type enzyme, the relation kcat/KNAD+ is 1.6times higher than in the wild type, resulting in an enzyme that prefers NAD+ over NADP+
G206D/R207I
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active only with NAD+; no activity with NADP+, highest reaction rate with cofactor NAD+ of all mutants tested; the mutant shows no activity with NADP+, when the coenzyme NAD+ is incubated with this double mutant, it reaches the highest kcat value, between 1.5 and 2times higher than the kcat of the wild type enzyme with NADP+, and between 3 and 4times higher than the kcat of the wild type with NAD+
G206D/R207I/R208N
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active only with NAD+; no activity with NADP+; the mutant shows no activity with NADP+
R207I
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less efficient with NAD+ or NADP+ than the wild-type enzyme; less efficient with NADP+ than the wild type; the mutant is less efficient with NADP+ than the wild type enzyme, shows an increase of 48times in Km value with NADP+ when compared with the wild type accompanied by a decrease in kcat, which clearly makes the R207I mutant less efficient in catalysis with NADP+, the R207I substitution also makes the enzyme less efficient with NAD+, with a decrease of 4 times in kcat/Km, this substitution also increases the Km for glucose
D38C
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crystallization of a range of binary and ternary complexes of the wild-type and a D38C mutant protein
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additional information
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Escherichia coli strain expressing both recombinant glucose 1-dehydrogenase and a glucose facilitator for uptake of unphosphorylated glucose shows a nine times higher initial alpha-pinene oxide formation rate corresponding to a sixfold higher yield of 20 mg per g cell dry weight after 1.5 h and to a sevenfold increased alpha-pinene oxide yield in the presence of glucose compared to glucose-free conditions; introduction of both, GLF and GlcDH, in P450-overexpressing Escherichia coli should enable the cell to carry out a straightforward intracellular cofactor regeneration driven by externally added glucose. For the generation of recombinant Escherichia coli strains carrying two plasmids, these are transformed successively. Firstly, pZY507glf is transformed into Escherichia coli BL21 (DE3). Subsequently, competent cells are prepared from a positive transformant, and then pETDUETbm-3qm glcdh is transformed.
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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method for determination of D-glucose- and D-galactose levels in glycoconjugates. The NAD(P)H produced from the enzymatic oxidation of the monosaccharides reacts with a CuSO4-bathocuproinedisulfonic acid reagent to produce a color complex absorbing maximally at 486 nm. With galactose dehydrogenase and glucose dehydrogenase serving as the model enzymes, reaction analysis gives a linear plot from 2.5 to 250 nmol of sugar. Method has been applicated to sugar released by acid hydrolysis from lactose, porcine submaxillary mucin and raffinose was quantified
industry
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Optimizing whole-cell biocatalysts by integrating a recombinant intracellular NADPH regeneration system through co-expression of a glucose facilitator from Zymomonas mobilis for uptake of unphosphorylated glucose and a NADP+-dependent glucose dehydrogenase from Bacillus megaterium that oxidizes glucose to gluconolactone.
synthesis