Information on EC 5.1.3.B12 - Agrobacterium tumefaciens D-psicose 3-epimerase

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 expected taxonomic range for this enzyme is: Agrobacterium tumefaciens

EC NUMBER
COMMENTARY hide
5.1.3.B12
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
Agrobacterium tumefaciens D-psicose 3-epimerase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
D-psicose = D-fructose
show the reaction diagram
-
-
-
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-fructose
D-psicose
show the reaction diagram
D-psicose
D-fructose
show the reaction diagram
D-sorbose
D-tagatose
show the reaction diagram
D-tagatose
D-sorbose
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
1 mM, 2.7fold increase in the epimerization rate from D-fructose to D-psicose
additional information
-
metal ion with octahedral coordination to two water molecules and four amino acid residues
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
-
1 mM, 98% inhibition
EDTA
-
activity of D-psicose 3-epimerase is 20% after the removal of metal ions by treatment with EDTA
Zn2+
-
1 mM, 96% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1 - 427
D-fructose
1 - 100
D-psicose
6 - 1640
D-tagatose
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.029 - 2068
D-fructose
23 - 2381
D-psicose
11 - 194
D-tagatose
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00029 - 85
D-fructose
117
0.44 - 205
D-psicose
2077
0.014 - 2.1
D-tagatose
2197
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33071
-
4 * 33071, MALDI-TOF MS
33093
-
4 * 33093, calculated from sequence
132000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of DPEase and its complex with the true substrate D-fructose. Diffraction-quality crystals of native and SeMet-enzyme are grown in 14–15% (w/v) PEG 1000, 0.1 M imidazole (pH 8.0), 0.2 M calcium acetate within five days to overall dimensions of 0.4 mm * 0.28 mm * 0.28 mm
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
250 min, stable. Half-life is 1265 min
60
-
half-life: 3.99 min
60.5
-
melting temperature of wild-type enzyme
63.6
-
melting temperature of S213C enzyme
64.8
-
melting temperature of I33L enzyme
68.1
-
melting temperature of S213C enzyme
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli. It has no significant sequence similarities to D-tagatose 3-epimerase. It is genetically different from other sugar epimerases
-
the gene AAL45544.1 from Agrobacterium tumefaciens str. C58 is modified by artificial synthesis for overexpression in Escherichial coli, expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A107I
-
7.7fold decrease in kcat/Km for D-fructose, no activity detected with D-tagatose
A107P
-
1.4fold decrease in kcat/Km for D-fructose, 6.6fold decrease in kcat/Km for D-tagatose
A107V
-
13.9fold decrease in kcat/Km for D-fructose, no activity detected with D-tagatose
E150Q
-
inactive mutant enzyme
E156D
-
mutant exhibits 63% of the wild-type activity
E244Q
-
inactive mutant enzyme
G67C
-
the variant shows remarkably decreased specific activity
I33E
-
the enzyme variant shows no activity
I33K
-
the enzyme variant shows no activity
I33L
-
increase of 5°C in the temperature for maximal enzyme activity, increase of 7.2-fold in the half-life at 50°C, and increase of 4.3°C in apparent melting temperature, respectively, compared with the wild-type enzyme
I33L/S213C
-
increase of 7.5°C in the temperature for maximal enzyme activity, increase of 29.9-fold in the half-life at 50°C, and increase of 7.6°C in apparent melting temperature, respectively, compared with the wild-type enzyme. After 8 or 16 days, the enzyme activity gradually decreases, and the conversion yields with and without borate are reduced to 22 and 9.6%, respectively, at 30 days. In contrast, the activity of the immobilized I33L/S213C variant with and without borate does not decrease during the operation time of 30 days
I33P
-
the enzyme variant shows no activity
I66A
-
1.4fold increase in kcat/Km for D-fructose, 150fold decrease in kcat/Km for D-tagatose
I66C
-
3fold decrease in kcat/Km for D-fructose, 29.2fold decrease in kcat/Km for D-tagatose
I66F
-
2.1fold decrease in kcat/Km for D-fructose, 23.3fold decrease in kcat/Km for D-tagatose
I66G
-
1.9fold decrease in kcat/Km for D-fructose, 87.5fold decrease in kcat/Km for D-tagatose
I66L
-
1.6fold increase in kcat/Km for D-fructose, 58,3fold decrease in kcat/Km for D-tagatose
I66V
-
1.9fold decrease in kcat/Km for D-fructose, 24fold decrease in kcat/Km for D-tagatose
I66W
-
1.4fold decrease in kcat/Km for D-fructose, 42fold decrease in kcat/Km for D-tagatose
I66Y
-
3fold decrease in kcat/Km for D-fructose, 26fold decrease in kcat/Km for D-tagatose
R215K
-
mutant exhibits 25% of the wild-type activity
S213C
-
increase of 2.5°C in the temperature for maximal enzyme activity, increase of 3.3-fold in the half-life at 50°C, and increase of 3.1°C in apparent melting temperature, respectively, compared with the wild-type enzyme. Immobilized wild-type enzyme with and without borate maintains activity for 8 days at a conversion yield of 70% (350 g/l psicose) and for 16 days at a conversion yield of 30% (150 g/l psicose), respectively
S213E
-
decrease of 7.2°C in half-life at 55°C
S213K
-
the variant shows no activity
S213M
-
decrease of 6.8°C in half-life at 55°C
S213P
-
decrease of 6°C in half-life at 55°C
S213T
-
decrease of 5°C in half-life at 55°C
S8T
-
the variant displays 29% of the wild-type activity
V96A
-
the variant displays 51% of the wild-type activity
W112A
-
mutant enzyme exhibits no detectable activity
W112F
-
mutant enzyme displays 19% of the wild type enzyme activity. kcat/Km for D-fructose is 100fold lower than wild-type value, kcat/Km fur D-psicose is 84fold lower than wild-type value
W112H
-
mutant enzyme displays 65% of the wild type enzyme activity
W112Y
-
mutant enzyme displays 54% of the wild type enzyme activity
E150Q
-
inactive mutant enzyme
-
E156D
-
mutant exhibits 63% of the wild-type activity
-
E244Q
-
inactive mutant enzyme
-
I33L
-
increase of 5°C in the temperature for maximal enzyme activity, increase of 7.2-fold in the half-life at 50°C, and increase of 4.3°C in apparent melting temperature, respectively, compared with the wild-type enzyme
-
I33L/S213C
-
increase of 7.5°C in the temperature for maximal enzyme activity, increase of 29.9-fold in the half-life at 50°C, and increase of 7.6°C in apparent melting temperature, respectively, compared with the wild-type enzyme. After 8 or 16 days, the enzyme activity gradually decreases, and the conversion yields with and without borate are reduced to 22 and 9.6%, respectively, at 30 days. In contrast, the activity of the immobilized I33L/S213C variant with and without borate does not decrease during the operation time of 30 days
-
I66A
-
1.4fold increase in kcat/Km for D-fructose, 150fold decrease in kcat/Km for D-tagatose
-
I66C
-
3fold decrease in kcat/Km for D-fructose, 29.2fold decrease in kcat/Km for D-tagatose
-
I66G
-
1.9fold decrease in kcat/Km for D-fructose, 87.5fold decrease in kcat/Km for D-tagatose
-
I66L
-
1.6fold increase in kcat/Km for D-fructose, 58,3fold decrease in kcat/Km for D-tagatose
-
I66V
-
1.9fold decrease in kcat/Km for D-fructose, 24fold decrease in kcat/Km for D-tagatose
-
R215K
-
mutant exhibits 25% of the wild-type activity
-
S213C
-
increase of 2.5°C in the temperature for maximal enzyme activity, increase of 3.3-fold in the half-life at 50°C, and increase of 3.1°C in apparent melting temperature, respectively, compared with the wild-type enzyme. Immobilized wild-type enzyme with and without borate maintains activity for 8 days at a conversion yield of 70% (350 g/l psicose) and for 16 days at a conversion yield of 30% (150 g/l psicose), respectively
-
S213E
-
decrease of 7.2°C in half-life at 55°C
-
S213K
-
the variant shows no activity
-
W112F
-
mutant enzyme displays 19% of the wild type enzyme activity. kcat/Km for D-fructose is 100fold lower than wild-type value, kcat/Km fur D-psicose is 84fold lower than wild-type value
-
W112H
-
mutant enzyme displays 65% of the wild type enzyme activity
-
W112Y
-
mutant enzyme displays 54% of the wild type enzyme activity
-