Information on EC 1.14.13.162 - 2,5-diketocamphane 1,2-monooxygenase

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The expected taxonomic range for this enzyme is: Pseudomonas putida

EC NUMBER
COMMENTARY hide
1.14.13.162
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RECOMMENDED NAME
GeneOntology No.
2,5-diketocamphane 1,2-monooxygenase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(+)-bornane-2,5-dione + O2 + NADH + H+ = (+)-5-oxo-1,2-campholide + NAD+ + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Bayer-Villiger reaction
oxidation
-
-
-
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redox reaction
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-
-
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reduction
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
(+)-camphor degradation
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SYSTEMATIC NAME
IUBMB Comments
(+)-bornane-2,5-dione,NADH:oxygen oxidoreductase (1,2-lactonizing)
A flavoprotein (FMN) which requires Fe2+. A Baeyer-Villiger monooxygenase isolated from camphor-grown strains of Pseudomonas putida and encoded on the cam plasmid. Involved in the degradation of (+)-camphor. Requires a dedicated NADH-FMN reductase [cf. EC 1.5.1.42, FMN reductase (NADH)] [1-3]. Can accept several bicyclic ketones including (+)- and (-)-camphor [6] and adamantanone [4]. The product spontaneously converts to [(1R)-2,2,3-trimethyl-5-oxocyclopent-3-enyl]acetate.
CAS REGISTRY NUMBER
COMMENTARY hide
37256-81-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain C1 ATCC 17453
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Manually annotated by BRENDA team
strain NCIMB 10007
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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
(+)-bornane-2,5-dione + NADH + O2
3,4,4-trimethyl-5-carboxy-methyl-DELTA2-cyclopentenone + NAD+ + H2O
show the reaction diagram
(+)-bornane-2,5-dione + NADH + O2
?
show the reaction diagram
(+)-bornanone + NADH + O2
1,2-campholide + NAD+ + H2O
show the reaction diagram
1,3,3-trimethyl-2-oxabicyclo-(2,2,2)octane + NADH + O2
1,6,6-trimethyl-2,7-dioxa(3,2,2)bicyclononan-3-one + NAD+ + H2O
show the reaction diagram
6-oxocineole + NADH + O2
3-(1-hydroxy-1-methylethyl)-6-oxoheptanoic acid + NAD+ + H2O
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
(+)-bornane-2,5-dione + NADH + O2
?
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2'-bipyridine
DTNB
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mild inhibition, dehydrogenase
H2O2
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mild inhibition, dehydrogenase
KI
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dehydrogenase
methylene blue
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dehydrogenase
NEM
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partial inhibition, dehydrogenase
p-hydroxymercuribenzoate
Sodium arsenite
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mild inhibition, dehydrogenase
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dichlorophenolindophenol
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activation, enhances NADH-oxidation
additional information
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boiled cell-free extract, activation, replaces FMN in the assay
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 0.019
FAD
0.0004 - 0.003
FMN
0.1 - 0.21
NADH
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
650
FAD
Pseudomonas putida
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component E2
20 - 317
NADH
additional information
additional information
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.335
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substrate (+)-camphor
0.43
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substrate (+)-camphor, purified, reconstituted multimeric enzyme complex
0.81
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substrate diketocamphane, purified enzyme
1.67
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component E2, lactonizing
3.56
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oxygenating component
10 - 20
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component E3, ketolactonizing
24
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component E2
38.3
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component E1, FMN-reductase
417
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purified component E1 NADH-dehydrogenase
420
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purified component E1, FMN-coupled NADH-dehydrogenase
additional information
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no enzyme assay possible in crude extract due to competition with other enzymes for NADH
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
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NADH-dehydrogenase
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8.5
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80% of activity maximum at pH 6.5 and pH 8.5
7 - 8
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70% of activity maximum at pH 7 and pH 8, NADH-dehydrogenase
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37000
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2 * 37000, oxygenating component E1, SDS-PAGE
39000 - 40000
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oxygenating component, gel filtration
40000
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component E1, analytical centrifugation
44000
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about, oxygenating component, native PAGE
76000
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oxygenating component E1, equilibrium ultracentrifugation
80000
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component E2, analytical centrifugation
120000
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal clusters, grown in 0.2 M CaCl2, 0.1 M HEPES and 30% PEG 400, X-ray analysis
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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10 min, component E2 retains full activity, component E3 loses 50%, component E3 loses 90% in the presence of substrate or component E1
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15°C, in 0.05 M Tris/HCl or potassium phosphate buffer, pH 7.2, the dehydrogenase is stable for months
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-20°C, ammonium sulfate precipitates as frozen pastes stable over a long period of time
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0°C, in buffer, the dehydrogenase is stable for a week, in very dilute solutions, the dehydrogenase is stable without protective agents
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
both components
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component E1, FMN-reductase
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component E1, FMN-reductase; component E2, lactonizing enzyme
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separation of components E1, E2, E3
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Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
inactive enzyme after dialysis can be reactivated by Fe2+ at 0.1 mM, Fe3+, Co2+, Zn2+, Mn2+, Mg2+, and Cu2+ are not usefull for reactivation
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis