Information on EC 1.1.99.28 - glucose-fructose oxidoreductase

Word Map on EC 1.1.99.28
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.99.28
-
RECOMMENDED NAME
GeneOntology No.
glucose-fructose oxidoreductase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-Glucose + D-fructose = D-gluconolactone + D-glucitol
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
sorbitol biosynthesis II
-
-
SYSTEMATIC NAME
IUBMB Comments
D-Glucose:D-fructose oxidoreductase
D-mannose, D-xylose, D-galactose, 2-deoxy-D-glucose and L-arabinose will function as aldose substrates, but with low affinities. The ketose substrate must be in the open-chain form. The apparent affinity for fructose is low, because little of the fructose substrate is in the open-chain form. Xylulose and glycerone (dihydroxyacetone) will replace fructose, but they are poor substrates. The enzyme from Zymomonas mobilis contains tightly bound NADP+.
CAS REGISTRY NUMBER
COMMENTARY hide
94949-35-6
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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 + D-fructose
?
show the reaction diagram
D-Galactose + D-fructose
?
show the reaction diagram
-
at 8% of the activity relative to D-glucose
-
-
-
D-Glucose + D-fructose
?
show the reaction diagram
D-glucose + D-fructose
D-glucono-1,5-lactone + D-glucitol
show the reaction diagram
-
-
-
-
?
D-glucose + D-fructose
D-glucono-1,5-lactone + D-sorbitol
show the reaction diagram
D-glucose + D-fructose
sorbitol + D-glucono-1,5-lactone
show the reaction diagram
-
-
-
-
?
D-glucose + D-fructose + H2O
D-glucono-1,5-lactone + D-sorbitol
show the reaction diagram
D-glucose + D-xylose
D-gluconolactone + D-xylitol
show the reaction diagram
-
-
-
?
D-Glucose + D-xylulose
?
show the reaction diagram
-
at 7% of the activity relative to fructose
-
-
-
D-Glucose + dihydroxyacetone
?
show the reaction diagram
-
at 6% of the activity relative to fructose
-
-
-
D-Glucose + L-sorbose
?
show the reaction diagram
-
at 0.5% of the activity relative to fructose
-
-
-
D-Mannose + D-fructose
?
show the reaction diagram
-
at 12% of the activity relative to D-glucose
-
-
-
D-Xylose + D-fructose
?
show the reaction diagram
-
at 8% of the activity relative to D-glucose
-
-
-
DL-Glyceraldehyde + D-fructose
?
show the reaction diagram
-
at 1.5% of the activity relative to D-glucose
-
-
-
L-Arabinose + D-fructose
?
show the reaction diagram
-
at 3% of the activity relative to D-glucose
-
-
-
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-Glucose + D-fructose
?
show the reaction diagram
D-glucose + D-fructose
D-glucono-1,5-lactone + D-glucitol
show the reaction diagram
-
-
-
-
?
D-glucose + D-fructose
D-glucono-1,5-lactone + D-sorbitol
show the reaction diagram
D-glucose + D-fructose + H2O
D-glucono-1,5-lactone + D-sorbitol
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADH
-
a single site mutation S116D alters the enzyme which in the wild type situation contains NAD(P)+ as nondissociable redox cofactor reacting in a ping-pong type mechanism to a dehydrogenase with dissociable NAD(P)+ as cosubstrate and a sequential reaction type
NADP+
NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Aldonolactone
-
the interaction of the enzyme with the aldonolactone product triggers a sequential process that affects the protein structure conformationally and chemically and results in an irreversible loss of activity
-
ethanol
-
16% inhibition at 2 M, 48% inhibition at 4.7 M
sorbitol
-
0.8 M, 27% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Triton X-100
-
activates
Tween
-
activates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
400
fructose
-
-
20
glucose
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
200
D-fructose
0.8
gluconolactone
additional information
additional information
Zymomonas mobilis
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.6
-
raw extract, at pH 6.3 and 25C
1.102
for D-xylitol
68.39
for D-sorbitol
72
-
purified enzyme, at pH 6.3 and 25C
245 - 310
-
-
additional information
-
kinetic parameters of the glucose dehydrogenase reaction catalyzed by mutant enzymes
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
-
-
7.8 - 8.2
-
higher enzymatic activities in immobilized permeabilized cells are obtained at pH 7.8 and 8.2 which are 80% higher than at pH 6.4
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.1
-
pH 5.0: about 35% of maximal activity, pH 7.1: about 30% of maximal activity
5.6 - 7.4
-
at pH 5.6 and 7.4 there are still 65% of enzyme activity remaining
7.7 - 8.7
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47 - 50
-
higher enzymatic activities in immobilized permeabilized cells are obtained at 47 and 50C which are 80% higher than at 39C, respectively
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
44 - 52
-
an increase of approximately 35% in the enzymatic activity in immobilized permeabilized cells is observed between 44 and 52C
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
translocated across the membrane in fully folded form. A twin-arginine motif in the signal peptide directs it to a Sec-independent pathway by which it is translocated, in fully folded form, into the periplasm where it functions to produce sorbitol for osmoprotection
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
Zymomonas mobilis subsp. mobilis (strain ATCC 31821 / ZM4 / CP4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
170000
-
gel filtration at pH 5.0
300000
-
gel filtration at pH 7.0
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop method, crystal structure of oxidized preGFOR R30K/R31K, in complex with succinate (preGFOR(succ)) and with glycerol (PreGFOR(gll)), at 2.2 A and 2.05 A resolution, respectively, and of reduced preGFOR R30K/R31K, after incubation with glucose (preGFOR(Glu)) and with sorbitol (preGFOR(sorb)) at 2.5 A and 2.6 A resolution. In all four crystal structures, the signal peptide is disordered, implying a flexibility that may be important for its interaction with the translocation apparatus. The crystal structures show that the mature enzyme portion of preGFOR is identical to native GFOR, in structure and cofactor binding
-
structure at 2.7 A
-
vapour diffusion in hanging drops, crystal structure of the NAD+ complex of a truncated form of the enzyme, GFORDELTA1-22/S64D, in which the first 22 residues of the N-terminal arm of the mature protein have been deleted, structure refined at 2.7 A resolution shows that the truncated form of the enzyme forms a dimer and implies that the N-terminal arm is essential for tetramer formation by wild-type GFOR
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.8
-
30C, pH does not affect the stability
10911
7
-
40C, fast inactivation above
10911
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
stable for 2 weeks
35
-
52 h, 10% loss of activity
39 - 45
-
when measured on its natural substrates (D-fructose and D-glucose), high enzyme thermal stability at 39 and 43C is observed, with remaining activities close to 100% of the initial, whereas at 45C, 91% of the activity is maintained
45
-
half-life: 55 h
50
-
the enzymatic activity in non-immobilized permeabilized cells is almost completely depleted at 50C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme activity loss in pineapple juice is even stronger (18%) than in 0.5 M sugar solution
-
guanidinium hydrochloride inactivates by induction of structural transitions that are comparable to that observed during substrate turnover. This leads to time-dependent formation of high-order associates and consequently inactivation
-
in absence of substrates or in presence of only one substrate, either fructose or glucose, the enzyme is fully stable
-
inactivation during substrate conversion
-
inactivation during substrate turnover. The process of inactivation is triggered by structural transitions that are induced by the lactone product and involves aggregation as the ultimate cause of irreversible inactivation
-
the interaction of the enzyme with the aldonolactone product triggers a sequential process that affects the protein structure conformationally and chemically and, ultimately, results in an irreversible loss of activity
-
thiol reagents stabilize. Dithiothreitol is the most efficient, 5-15 mM
-
urea, 1.0 M, prevents the formation of high-order associates and increases the half-life under operational conditions 10fold
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
mutant enzyme DELTA32-46
-
mutant enzyme S116D
-
Sepharose CL-4B column chromatography and Source S column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
expression in Escherichia coli; NADP+ is required for the enzyme to fold into its native conformation and its absence from the Escherichia coli periplasm is responsible for failure to form a stable periplasmic protein
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
DELTA1-22/S64D
-
S64D mutation converts the strict NADP+ spoecificity of wild-type GFOR to a dual NADP+/NAD+ specificity
DELTA32-46
-
the mutant enzyme DELTA32-46 is a protein that is no longer processed but shows full enzymatic activity and has the cofactor firmly bound. The mutant enzyme DELTA2-20 or a mutant enzyme with an exchange of the entire signal sequence with the signal sequence of gluconolactonase of Zymomonas mobilis leads to an active and processed protein
K121A
-
mutant enzyme is not processed within 60 min
K123A
-
mutant enzyme shows processing behavior comparable to wild-type enzyme
S116D/K121A/K123Q/I124K
-
significantly retarded processing kinetics with residual unprocessed form being detectable even after 60 min
Y269F
essential acid-base catalyst, involved in substrate binding, activity completely abolished
additional information
-
mutant enzymes with deletions in the signal peptide are enzymatically active and contain tightly bound NADP(H). Mutant enzymes with a complete deletion of the signal peptide are produced as cytosolic enzymes
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (458 entries)
Please use the Sequence Search for a certain query.