Information on EC 1.1.99.35 - soluble quinoprotein glucose dehydrogenase

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The expected taxonomic range for this enzyme is: Acinetobacter calcoaceticus

EC NUMBER
COMMENTARY hide
1.1.99.35
-
RECOMMENDED NAME
GeneOntology No.
soluble quinoprotein glucose dehydrogenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-glucose + acceptor = D-glucono-1,5-lactone + reduced acceptor
show the reaction diagram
SYSTEMATIC NAME
IUBMB Comments
D-glucose:acceptor oxidoreductase
Soluble periplasmic enzyme containing PQQ as prosthetic group, bound to a calcium ion. Electron acceptor is not known. It is assayed with Wurster's Blue or phenazine methosulfate. It has negligible sequence or structure similarity to other quinoproteins. It catalyses an exceptionally high rate of oxidation of a wide range of aldose sugars, including D-glucose, galactose, arabinose and xylose, and also the disaccharides lactose, cellobiose and maltose. It has been described only in Acinetobacter calcoaceticus.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-deoxy-D-glucose + oxidized 2,6-dichlorophenolindophenol
2-deoxy-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
3-O-methyl-D-glucose + oxidized 2,6-dichlorophenolindophenol
3-O-methyl-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
cellobiose + oxidized 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
D-allose + oxidized 2,6-dichlorophenolindophenol
D-allono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-fucose + oxidized 2,6-dichlorophenolindophenol
D-fucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
D-galactose + oxidized 2,6-dichlorophenolindophenol
D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-galactose + oxidized N-methylphenazonium methyl sulfate
D-galactono-1,5-lactone + reduced N-methylphenazonium methyl sulfate
show the reaction diagram
-
-
-
-
?
D-glucose + acceptor
D-glucono-1,5-lactone + reduced acceptor
show the reaction diagram
-
-
-
-
?
D-glucose + oxidized 2,6-dichlorophenol-indophenol
D-glucono-1,5-lactone + reduced 2,6-dichlorophenol-indophenol
show the reaction diagram
-
-
-
-
?
D-glucose + oxidized 2,6-dichlorophenolindophenol
D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-glucose + oxidized N-methylphenazonium methyl sulfate
D-glucono-1,5-lactone + reduced N-methylphenazonium methyl sulfate
show the reaction diagram
-
-
-
-
?
D-lyxose + oxidized 2,6-dichlorophenolindophenol
D-lyxono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
D-mannose + oxidized 2,6-dichlorophenolindophenol
D-mannono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-ribose + oxidized 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
D-xylose + oxidized 2,6-dichlorophenolindophenol
D-xylono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-xylose + oxidized N-methylphenazonium methyl sulfate
D-xylono-1,5-lactone + reduced N-methylphenazonium methyl sulfate
show the reaction diagram
-
-
-
-
?
L-arabinose + oxidized 2,6-dichlorophenolindophenol
L-arabinono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
lactose + oxidized 2,6-dichlorophenol-indophenol
? + reduced 2,6-dichlorophenol-indophenol
show the reaction diagram
-
-
-
-
?
lactose + oxidized 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
lactose + oxidized 2,6-dichlorophenolindophenol
lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
lactose + oxidized N-methylphenazonium methyl sulfate
4-O-beta-D-galactopyranosyl-D-glucono-1,5-lactone + reduced N-methylphenazonium methyl sulfate
show the reaction diagram
-
-
-
-
?
maltose + acceptor
maltono-1,5-lactone + reduced acceptor
show the reaction diagram
-
-
-
-
?
maltose + oxidized 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
maltose + oxidized 2,6-dichlorophenolindophenol
maltono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
melibiose + oxidized 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
-
-
-
?
additional information
?
-
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-galactose + oxidized 2,6-dichlorophenolindophenol
D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-glucose + acceptor
D-glucono-1,5-lactone + reduced acceptor
show the reaction diagram
-
-
-
-
?
D-glucose + oxidized 2,6-dichlorophenolindophenol
D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-mannose + oxidized 2,6-dichlorophenolindophenol
D-mannono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
about 11% activity compared to D-glucose
-
-
?
D-xylose + oxidized 2,6-dichlorophenolindophenol
D-xylono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
about 9% activity compared to D-glucose
-
-
?
lactose + oxidized 2,6-dichlorophenolindophenol
lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
about 50% activity compared to D-glucose
-
-
?
maltose + oxidized 2,6-dichlorophenolindophenol
maltono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
-
about 70% activity compared to D-glucose
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyrroloquinoline quinone
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
or Ca2+, Sr2+, or Mn2+, required for binding of cofactor PQQ in soluble isoform sGDH. Mg2+, or Ca2+, Zn2+, or Sr2+, required for binding of cofactor PQQ in membrane-bound isoform mGDH
Mg2+
-
or Ca2+, Zn2+, or Sr2+, required for binding of cofactor PQQ in membrane-bound isoform mGDH. With Mg2+, 115% of the activity with Ca2+
Mn2+
-
or Ca2+, Sr2+, or Cd2+, required for binding of cofactor PQQ in soluble isoform sGDH. Mg2+, or Ca2+, Zn2+, or Sr2+, required for binding of cofactor PQQ in membrane-bound isoform mGDH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
methylhydrazine
-
competitive inhibitor
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
88 - 90
2-deoxy-D-glucose
2 - 46
3-O-methyl-D-glucose
21 - 38.7
D-Allose
2.7 - 9
D-galactose
2 - 64
D-glucose
14.3 - 34
D-lactose
14.3 - 46.5
D-maltose
22
D-mannose
-
mutant E277K, pH 7.0; wild-type, pH 7.0
7.5 - 14.3
D-xylose
17.5 - 20
lactose
13 - 14
maltose
additional information
additional information
-
under conditions that avoid its masking by sugar-substrate inhibition as much as possible, negative kinetic cooperativity with respect to sugar substrate oxidation is observed. The cooperativity effect dramatically changes the performance of soluble GDH, as reflected by the V2 and K2 values for glucose in phosphate buffer being about 10-fold and 100-fold higher than the V1 and K1 values, respectively. Substituting the Ca2+ involved in activation of pyrroloquinoline quinone in soluble GDH by Sr2+ affects the cooperativity effect but not the two turnover rates of the hybrid enzyme for glucose
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
331 - 1063
2-deoxy-D-glucose
1064 - 3198
3-O-methyl-D-glucose
949 - 4563
D-Allose
69 - 630
D-galactose
1399 - 3860
D-glucose
201 - 1659
D-lactose
588 - 1930
D-maltose
267 - 861
D-mannose
669 - 1795
D-xylose
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4 - 12
2-deoxy-D-glucose
531
31 - 118
3-O-methyl-D-glucose
1525
1080
cellobiose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
82
25 - 1470
D-Allose
945
440
D-fucose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
1256
18 - 240
D-galactose
71
114 - 2100
D-glucose
35
7 - 88
D-lactose
3242
0.24
D-Lyxose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
1589
22 - 74
D-maltose
1168
12 - 50
D-mannose
216
11
D-ribose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
292
45 - 239
D-xylose
115
1450
L-arabinose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
206
790 - 960
lactose
114
800
maltose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
86
11
melibiose
Acinetobacter calcoaceticus
-
stopped-flow measurement of reductive half-reaction, 15C
606
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
92 - 156
D-glucose
61
lactose
-
pH 7.0, 20C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
515
-
crude cell lysate, at pH 7.0 and 25C
2977
-
wild-type, pH 7.0, at 50 mM D-glucose
3313
-
mutant S231K, pH 7.0, at 100 mM D-glucose
5811
-
after 11.3fold purification, at pH 7.0 and 25C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
pH optimum determined for glucose oxidation appears to be 7.0, implying that reoxidation of sGDHred is rate-limiting with those electron acceptors displaying a different value under steady-state conditions
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
2 * 50000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
study on heterodimeric PQQGDH-B composed of native wild-type and inactive mutant H168Q subunits
homodimer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
the secreted PQQGDH-B in Pichia pastoris is glycosylated but shows similar enzymatic properties as compared with those of recombinant PQQGDH-B produced in Escherichia coli
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 1.72 A resolution. The s-GDH monomer has a beta-propeller fold consisting of six four-stranded anti-parallel beta-sheets aligned around a pseudo 6-fold symmetry axis. The enzyme binds three calcium ions per monomer, two of which are located in the dimer interface. The third is bound in the putative active site, where it may bind and functionalize the pyrroloquinoline quinone cofactor
-
diffraction to beyond 2.1 A resolution, space group P21
-
strucuture of soluble isoform sGDH with the cofactor at 2.2 A resolution, and of its complex with reduced cofactor and D-glucose at 1.9 A resolution. Evidence for a mechanism comprisding general base-catalyzed hydride transfer
-
ternary complex of s-GDH with PQQ and methylhydrazine, at 1.5 A resolution. Formation of a covalent PQQ adduct in the active-site. The C5 carbonyl group of the cofactor is the most reactive moiety of PQQ. The binding of the cofactor to s-GDH is predominantly governed by polar interactions. The C2, C7, and C9 carboxyl groups of PQQ form salt bridges with Arg408, Lys377, and Arg406, respectively. The ortho-quinone O4 and O5 atoms are bound by Asn229 and Arg228, respectively. The N6, O7A, and O5 atoms of PQQ are ligands for the active-site calcium ion. The other calcium ligands are provided by the two main chain carbonyl oxygen atoms of Gly-247 and Pro-248 and two watermolecules
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
53.9
-
wild-type, 50% loss of initial activity within in 10 min
56.5
-
mutant T416V/T417V, 50% loss of initial activity within in 10 min
57.5
-
mutant D340F/Y418I, 50% loss of initial activity within in 10 min
57.7
-
mutant D340F/Y418F, 50% loss of initial activity within in 10 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme nonspecifically immobilized on carbon cryogel electrodes retains its enzymatic activity for D-glucose and maltose oxidation at pH 7.2 and 37C
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA affinity column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
-
expression in Klebsiella pneumoniae
-
expression in Pichia pastoris
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D275E
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 6%, D-mannose 11%, D-allose 44%, 3-O-methyl-D-glucose 45%, D-galactose 12%, D-xylose 7%, D-lactose 52%, D-maltose 44%, respectively
D276E
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 13%, D-mannose 19%, D-allose 54%, 3-O-methyl-D-glucose 57%, D-galactose 26%, D-xylose 20%, D-lactose 75%, D-maltose 66%, respectively
D340F/Y418F
-
increase in temperature stability
D340F/Y418I
-
increase in temperature stability
D448N
-
substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 72%, D-allose 39%, D-galactose 14%, D-maltose 36%, D-lactose 48%, respectively
D456N
-
substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 78%, D-allose 43%, D-galactose 16%, D-maltose 41%, D-lactose 59%, respectively
DD457N
-
substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 80%, D-allose 46%, D-galactose 11%, D-maltose 32%, D-lactose 43%, respectively
E277A
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 11%, D-mannose 33%, D-allose 142%, 3-O-methyl-D-glucose 91%, D-galactose 46%, D-xylose 23%, D-lactose 83%, D-maltose 39%, respectively
E277D
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 9%, D-mannose 19%, D-allose 68%, 3-O-methyl-D-glucose 58%, D-galactose 21%, D-xylose 15%, D-lactose 87%, D-maltose 73%, respectively
E277G
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 15%, D-mannose 38%, D-allose 189%, 3-O-methyl-D-glucose 113%, D-galactose 59%, D-xylose 28%, D-lactose 100%, D-maltose 52%, respectively
E277H
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 3%, D-mannose 11%, D-allose 67%, 3-O-methyl-D-glucose 45%, D-galactose 12%, D-xylose 9%, D-lactose 57%, D-maltose 39%, respectively
E277K
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 7%, D-mannose 18%, D-allose 116%, 3-O-methyl-D-glucose 84%, D-galactose 28%, D-xylose 18%, D-lactose 79%, D-maltose 74%, respectively
E277N
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 8%, D-mannose 29%, D-allose 148%, 3-O-methyl-D-glucose 91%, D-galactose 43%, D-xylose 18%, D-lactose 86%, D-maltose 59%, respectively
E277Q
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 10%, D-mannose 31%, D-allose 134%, 3-O-methyl-D-glucose 80%, D-galactose 35%, D-xylose 25%, D-lactose 68%, D-maltose 54%, respectively
E277V
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 8%, D-mannose 29%, D-allose 150%, 3-O-methyl-D-glucose 101%, D-galactose 25%, D-xylose 19%, D-lactose 114%, D-maltose 65%, respectively
I278F
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 4%, D-mannose 14%, D-allose 54%, 3-O-methyl-D-glucose 47%, D-galactose 15%, D-xylose 12%, D-lactose 64%, D-maltose 49%, respectively
N279H
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 6%, D-allose 49%, 3-O-methyl-D-glucose 50%, D-galactose 13%, D-xylose 8%, D-lactose 64%, D-maltose 61%, respectively
N452T
-
substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 59%, D-allose 33%, D-galactose 5%, D-maltose 30%, D-lactose 31%, respectively
N462H
-
substrate specificity: D-glucose 100% activity, 3-O-methyl-D-glucose 53%, D-allose 32%, D-galactose 2%, D-maltose 25%, D-lactose 31%, respectively
S231C
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 3%, D-mannose 8%, D-allose 46%, 3-O-methyl-D-glucose 76%, D-galactose 14%, D-xylose 8%, D-lactose 69%, D-maltose 69%, respectively
S231D
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 9%, D-allose 38%, 3-O-methyl-D-glucose 71%, D-galactose 12%, D-xylose 8%, D-lactose 54%, D-maltose 38%, respectively
S231H
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 2%, D-mannose 12%, D-allose 57%, 3-O-methyl-D-glucose 86%, D-galactose 17%, D-xylose 7%, D-lactose 56%, D-maltose 38%, respectively
S231K
-
increase in thermal stability. Substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 10%, D-allose 43%, 3-O-methyl-D-glucose 82%, D-galactose 15%, D-xylose 5%, D-lactose 59%, D-maltose 70%, respectively
S231L
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 6%, D-mannose 13%, D-allose 62%, 3-O-methyl-D-glucose 105%, D-galactose 20%, D-xylose 12%, D-lactose 73%, D-maltose 76%, respectively
S231M
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 9%, D-allose 43%, 3-O-methyl-D-glucose 80%, D-galactose 10%, D-xylose 8%, D-lactose 56%, D-maltose 41%, respectively
S231N
-
substrate specificity: D-glucose 100% activity, 2-deoxy-D-glucose 5%, D-mannose 12%, D-allose 61%, 3-O-methyl-D-glucose 109%, D-galactose 18%, D-xylose 15%, D-lactose 66%, D-maltose 51%, respectively
T416V/T417V
-
increase in temperature stability
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
energy production
-
a (PQQ)-GDH electrode is used as an anode to convert the chemical energy of D-glucose into electrical energy by oxidation of the substrate
synthesis