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Information on EC 1.1.99.32 - L-sorbose 1-dehydrogenase
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1.1.99.32 L-sorbose 1-dehydrogenaseIUBMB Comments
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.
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Word Map
- 1.1.99.32
-
2-keto-l-gulonic
- dehydrogenases
- gluconobacter
- vulgare
- ketogulonicigenium
- l-sorbosone
- oxydans
- d-sorbitol
-
homodimeric
-
quinoprotein
-
protrusion
-
l-ascorbic
-
quinone-dependent
-
bladed
- l-idonate
-
pyrroloquinoline
- choline
- synthesis
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
l-sorbose dehydrogenase, 
more
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
L-sorbose dehydrogenase
PQQ-dependent L-sorbose dehydrogenase
SDH
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-sorbose + acceptor = 1-dehydro-L-sorbose + reduced acceptor
-
-
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
redox reaction
reduction
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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.
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CAS REGISTRY NUMBER
COMMENTARY
133249-51-1
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
1,5-anhydro-D-glucitol + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
1,5-anhydro-D-glucitol + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
79% activity compared to L-sorbose
-
-
?
1,5-anhydro-D-glucitol + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
79% activity compared to L-sorbose
-
-
?
D-galactose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
1.4% activity compared to L-sorbose
-
-
?
D-galactose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
1.4% activity compared to L-sorbose
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
1.0% activity compared to L-sorbose
-
-
?
D-glucose + 2,6-dichlorophenolindophenol
? + reduced 2,6-dichlorophenolindophenol
-
1.0% activity compared to L-sorbose
-
-
?
L-sorbose + 2,6-dichloroindophenol
L-sorbosone + reduced 2,6-dichloroindophenol
-
-
-
-
?
L-sorbose + 2,6-dichloroindophenol
L-sorbosone + reduced 2,6-dichloroindophenol
-
-
-
-
?
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
100% activity
-
-
?
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
100% activity
-
-
?
L-sorbose + acceptor
L-sorbosone + reduced acceptor
-
-
-
-
?
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
-
-
-
-
?
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
-
-
-
-
?
L-sorbose + phenazine methosulfate
L-sorbosone + reduced phenazine methosulfate
-
-
-
-
?
L-sorbose + phenazine methosulfate
L-sorbosone + reduced phenazine methosulfate
-
-
-
-
?
additional information
?
-
-
enzyme is highly specific for L-sorbose. No substrates are glucose, mannitol, sorbitol, fructose, glycerol, adonitol, erythritol, gluconate, calcium idonate, mannose, maltose, xylose, ethanol, lactose, sucrose, galactose, cellobiose, inositol, arabinose, sodium 2-keto-L-gulonic acid
-
-
?
additional information
?
-
-
enzyme is highly specific for L-sorbose. No substrates are glucose, mannitol, sorbitol, fructose, glycerol, adonitol, erythritol, gluconate, calcium idonate, mannose, maltose, xylose, ethanol, lactose, sucrose, galactose, cellobiose, inositol, arabinose, sodium 2-keto-L-gulonic acid
-
-
?
additional information
?
-
-
less than 1% activity with D-fructose, D-cellobiose, 2-deoxy-D-glucose, methyl-beta-D-glucopyranoside, 3-O-methyl-D-glucose, D-glucose 6 phosphate, D-mannose, D-sucrose, D-arabinose, and D-raffinose. No activity with arabitol, D-xylose, and glycerol
-
-
?
additional information
?
-
-
less than 1% activity with D-fructose, D-cellobiose, 2-deoxy-D-glucose, methyl-beta-D-glucopyranoside, 3-O-methyl-D-glucose, D-glucose 6 phosphate, D-mannose, D-sucrose, D-arabinose, and D-raffinose. No activity with arabitol, D-xylose, and glycerol
-
-
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
L-sorbose + 2,6-dichlorophenolindophenol
L-sorbosone + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
L-sorbose + acceptor
L-sorbosone + reduced acceptor
-
-
-
-
?
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
-
-
-
-
?
L-sorbose + cytochrome c551
L-sorbosone + reduced cytochrome c551
-
-
-
-
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
L-sorbose dehydrogenase accelerative protein (SAP)
isolated from Bacillus megaterium
-
Triton X-100
-
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
-
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
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
100
L-sorbose
-
electron acceptor 2,6-dichloroindophenol, pH 7.0, 25°C
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
enzyme activity is induced by growth on L-sorbose or D-sorbitol as carbon source
-
-
brenda
enzyme activity is induced by growth on L-sorbose or D-sorbitol as carbon source
-
-
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). F7ZJ19_ROSLO
Roseobacter litoralis (strain ATCC 49566 / DSM 6996 / JCM 21268 / NBRC 15278 / OCh 149)
546
0
58696
TrEMBL
-
A0A1S7QKN4_RHIRD
551
0
60039
TrEMBL
-
A0A0E0UL85_SINMB
Sinorhizobium meliloti (strain BL225C)
531
0
57278
TrEMBL
-
A0A067Z809_GLUOY
531
0
57537
TrEMBL
-
A0A1S7S7U2_RHIRD
551
0
59983
TrEMBL
-
A0A0E1CMR2_KLEPN
317
0
34141
TrEMBL
-
F7ZFI5_ROSLO
Roseobacter litoralis (strain ATCC 49566 / DSM 6996 / JCM 21268 / NBRC 15278 / OCh 149)
534
0
57626
TrEMBL
-
A0A6N3LH57_9VIBR
540
0
58542
TrEMBL
-
A0A822V8Q3_RHIRD
551
0
59979
TrEMBL
-
A0A6N3LGC4_9VIBR
531
0
58064
TrEMBL
-
A0A1S7TFK3_AGRFH
528
0
57765
TrEMBL
-
I3XEL4_SINF2
Sinorhizobium fredii (strain USDA 257)
551
0
60044
TrEMBL
-
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
tetramer
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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
-
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 55
-
the enzyme is stable at temperatures of up to 30°C, and 82% of enzyme activity persists at 35°C. Enzyme activity is almost completely abrogated by a 10 min incubation at 55°C
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
rather stable in buffer containing 200 mM sorbose, pH 7.0, below 30°C
-
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography, HiTrap Q column chromatography, and Superdex 200 gel filtration
-
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
synthesis
-
the enzyme (SDH) can be used to directly produce 2-keto-L-gulonic acid from L-sorbose with high substrate/product specificity. A platform strain suitable for highthroughput screening of SDH is constructed
synthesis
Gluconobacter oxydans WSH-004
-
the enzyme (SDH) can be used to directly produce 2-keto-L-gulonic acid from L-sorbose with high substrate/product specificity. A platform strain suitable for highthroughput screening of SDH is constructed
-
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REF.
AUTHORS
TITLE
JOURNAL
VOL.