Information on EC 1.1.99.32 - L-sorbose 1-dehydrogenase

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The expected taxonomic range for this enzyme is: Alphaproteobacteria

EC NUMBER
COMMENTARY hide
1.1.99.32
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RECOMMENDED NAME
GeneOntology No.
L-sorbose 1-dehydrogenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-sorbose + acceptor = 1-dehydro-L-sorbose + reduced acceptor
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ascorbate metabolism
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L-ascorbate biosynthesis III
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2-keto-L-gulonate biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
L-sorbose:acceptor 1-oxidoreductase
The product, L-sorbosone, is an intermediate in bacterial 2-keto-L-gulonic-acid formation. The activity of this membrane-bound enzyme is stimulated by Fe(III) or Co2+ but is inhibited by Cu2+. The enzyme is highly specific for L-sorbose as other sugars, such as glucose, mannitol and sorbitol, are not substrates. Phenazine methosulfate and DCIP can act as artificial acceptors.
CAS REGISTRY NUMBER
COMMENTARY hide
133249-51-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain UV10, enzyme activity is induced by growth on L-sorbose or D-sorbitol as carbon source
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Manually annotated by BRENDA team
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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
1,5-anhydro-D-glucitol + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-galactose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
D-glucose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
L-sorbose + 2,6-dichloroindophenol
L-sorbosone + reduced 2,6-dichloroindophenol
show the reaction diagram
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
L-sorbose + acceptor
L-sorbosone + reduced acceptor
show the reaction diagram
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
show the reaction diagram
L-sorbose + phenazine methosulfate
L-sorbosone + reduced phenazine methosulfate
show the reaction diagram
additional information
?
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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
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
show the reaction diagram
L-sorbose + acceptor
L-sorbosone + reduced acceptor
show the reaction diagram
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyrroloquinoline quinone
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
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0.8 mM, 18% increase in activity
Fe3+
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0.4 mM, 21% increase in activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
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0.8 mM, complete loss of activity
EDTA
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5.2 mM, 13% inhibition
iodoacetate
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4.2 mM, 34% inhibition
Quinine
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4.2 mM, 30% inhibition
sodium fluoroacetate
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9.4 mM, 26% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
L-sorbose dehydrogenase accelerative protein (SAP)
isolated from Bacillus megaterium
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Triton X-100
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solubilization with Triton X-100 gives 10fold increase in activity compared to solubilization with Tween 80, differential solubilization with Tween 80 + Triton X-100 gives 2fold higher activity than Triton X-100 alone
Tween 80
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solubilization with Triton X-100 gives 10fold increase in activity compared to solubilization with Tween 80, differential solubilization with Tween 80 + Triton X-100 gives 2fold higher activity than Triton X-100 alone
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
97.5
1,5-anhydro-D-glucitol
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at pH 8.5 and 40C
62.4 - 100
L-sorbose
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14.1
1,5-anhydro-D-glucitol
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at pH 8.5 and 40C
11.2
L-sorbose
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at pH 8.5 and 40C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
1,5-anhydro-D-glucitol
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at pH 8.5 and 40C
0.18
L-sorbose
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at pH 8.5 and 40C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.897
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pH 7.0, 25C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
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no decrease in activity is observed in the pH range 5.5-8.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
58000
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4 * 58000, SDS-PAGE
117200
calculated from amino acid sequence
150000
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gel filtration
210000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 1.7 M ammonium sulfate, 5 mM pyrroloquinoline quinone, 5 mM CaCl2, and 0.1 M pH 7.4 HEPES
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
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unstable below
677427
6.5 - 8
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677427
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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stable up to
30 - 55
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the enzyme is stable at temperatures of up to 30C, and 82% of enzyme activity persists at 35C. Enzyme activity is almost completely abrogated by a 10 min incubation at 55C
46
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5 min, 50% loss of activity
65
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5 min, 90% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, purified enzyme, 2 days, 30% loss of activity
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rather stable in buffer containing 200 mM sorbose, pH 7.0, below 30C
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HisTrap column chromatography, and Superdex 200 gel filtration
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Ni-NTA column chromatography, HiTrap Q column chromatography, and Superdex 200 gel filtration
purification from membrane fraction
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TALON metal affinity resin column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli codon plus cells