Information on EC 3.7.1.11 - cyclohexane-1,2-dione hydrolase

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: Azoarcus

EC NUMBER
COMMENTARY hide
3.7.1.11
-
RECOMMENDED NAME
GeneOntology No.
cyclohexane-1,2-dione hydrolase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cyclohexane-1,2-dione + H2O = 6-oxohexanoate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Caprolactam degradation
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
cyclohexane-1,2-dione acylhydrolase (decyclizing)
Highly specific; does not act on cyclohexanone or cyclohexane-1,3-dione as substrate.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
cyclohexane-1,2-dione + H2O
6-oxohexanoate
show the reaction diagram
cyclohexane-1,2-dione + H2O
6-oxohexanoate + ?
show the reaction diagram
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
cyclohexane-1,2-dione + H2O
6-oxohexanoate
show the reaction diagram
cyclohexane-1,2-dione + H2O
6-oxohexanoate + ?
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
-
-
thiamine diphosphate
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
activity decreases significantly at this high ionic strength
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-methyl-2,4-pentane-diol
-
MPD, competitive inhibitor, shows selective binding to the open funnel of CDH and strong binding to the active site. It contacts besides GlyA399 the apolar side chains of PheA259, TrpA285, LeuA551, LeuA487 and LeuA563
Cyclohexane-1,2-dione
-
substrate inhibition above 0.1 mM
NaCl
-
inhibitory above 50 mM due to binding of the chloride anion in close neighborhood to the thiamine diphosphate cofactor
additional information
-
activity decreases significantly at this high ionic strength
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
-
dependent on, one molecule ThDP per enzyme monomer
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0133
Cyclohexane-1,2-dione
-
pH 8.0, 37C
additional information
additional information
-
CDH follows Michaelis-Menten kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.6
Cyclohexane-1,2-dione
Azoarcus sp.
-
pH 8.0, 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0012
Cyclohexane-1,2-dione
Azoarcus sp.
-
pH 8.0, 37C
10620
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.677
-
purified enzyme, pH 8.0, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 30
-
assay at
30
-
assay at
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
105000
-
gel filtration
120000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis of CDH in complex with substrate cyclohexane-1,2-dione, PDB ID 2PGN
-
purified CDH from strain 22Lin, hanging drop vapour diffusion method, equal volumes of protein storage and reservoir solution, the latter composed of 60% MPD, 20 mM Na+ acetate and 200 mM NaCl, 25C, a second cubic crystal form is obtained at 20C with a reservoir solution containing 17-18% w/v PEG 8000, 100 mM Na+ acetate, 10 mM MnCl2, and 2% w/v isopropanol in 0.1 M HEPES buffer, pH 7.5, X-ray diffraction structure determination and analysis at 1.26 A resolution, modeling
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native CDH 6.6fold by two different steps of anion exchange chromatography, followed by gel filtration
-
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant expression of C-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H28A
-
site-directed mutagenesis, the mutant enzyme is much less able to catalyze the C-C bond formation as the wild-type enzyme, while the ability for C-C bond cleavage is still intact, the H28A variant shows an 8fold decrease in the formation of (R)-phenylacetylcarbinol (12%), but 1,2-diketone cleavage is nearly unaffected (78% conversion)
H28A/N484A
-
site-directed mutagenesis, the double mutant catalyzes the addition of pyruvate to cyclohexane-1,2-dione, resulting in the formation of a tertiary alcohol, variant H28A/N484A shows acceptable formation of (R)-phenylacetylcarbinol (73%), but conversion toward the cleavage product is decreased by a factor of five (17% conversion), the mutant is also active with 1,2-diketone in contrast to the wild-type enzyme, mutant substrate specificity amd enantioselectivity, overview
H76A
-
site-directed mutagenesis, inactive mutant
H76A/Q116A
-
site-directed mutagenesis, inactive mutant
Q116A
-
site-directed mutagenesis, inactive mutant
H28A
-
site-directed mutagenesis, the mutant enzyme is much less able to catalyze the C-C bond formation as the wild-type enzyme, while the ability for C-C bond cleavage is still intact, the H28A variant shows an 8fold decrease in the formation of (R)-phenylacetylcarbinol (12%), but 1,2-diketone cleavage is nearly unaffected (78% conversion)
-
H28A/N484A
-
site-directed mutagenesis, the double mutant catalyzes the addition of pyruvate to cyclohexane-1,2-dione, resulting in the formation of a tertiary alcohol, variant H28A/N484A shows acceptable formation of (R)-phenylacetylcarbinol (73%), but conversion toward the cleavage product is decreased by a factor of five (17% conversion), the mutant is also active with 1,2-diketone in contrast to the wild-type enzyme, mutant substrate specificity amd enantioselectivity, overview
-
H76A
-
site-directed mutagenesis, inactive mutant
-
Q116A
-
site-directed mutagenesis, inactive mutant
-
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
synthesis