Information on EC 6.2.1.14 - 6-carboxyhexanoate-CoA ligase

for references in articles please use BRENDA:EC6.2.1.14
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
6.2.1.14
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RECOMMENDED NAME
GeneOntology No.
6-carboxyhexanoate-CoA ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + 6-carboxyhexanoate + CoA = AMP + diphosphate + 6-carboxyhexanoyl-CoA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acid-thiol ligation
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
8-amino-7-oxononanoate biosynthesis III
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Biotin metabolism
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
6-carboxyhexanoate:CoA ligase (AMP-forming)
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CAS REGISTRY NUMBER
COMMENTARY hide
55467-50-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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UniProt
Manually annotated by BRENDA team
Bacillus amyloliquefaciens ATCC 23350 / DSM 7 / BCRC 11601 / NBRC 15535 / NRRL B-14393
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UniProt
Manually annotated by BRENDA team
weak activity
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
strain AKU 0007
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Manually annotated by BRENDA team
strain AKU 0022, strain IFO 3317, strain IFO 12010
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Lavandula vera
biotin-overproducing cell line
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
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
ADP + heptanedioate + CoA
AMP + phosphate + heptanedioyl-CoA
show the reaction diagram
ATP + 6-carboxyhexanoate + CoA
AMP + diphosphate + 6-carboxyhexanoyl-CoA
show the reaction diagram
ATP + adipate + CoA
AMP + diphosphate + adipyl-CoA
show the reaction diagram
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-
-
-
?
ATP + azelate + CoA
AMP + diphosphate + azelayl-CoA
show the reaction diagram
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-
-
-
?
ATP + glutarate + CoA
AMP + diphosphate + glutaryl-CoA
show the reaction diagram
ATP + heptanedioate + CoA
?
show the reaction diagram
ATP + heptanedioate + CoA
AMP + diphosphate + heptanedioyl-CoA
show the reaction diagram
ATP + heptanoate + CoA
AMP + diphosphate + heptanoyl-CoA
show the reaction diagram
ATP + hexanedioate + CoA
AMP + diphosphate + hexanedioyl-CoA
show the reaction diagram
ATP + nonanedioate + CoA
AMP + diphosphate + nonanedioyl-CoA
show the reaction diagram
ATP + octanoate + CoA
AMP + diphosphate + octanoyl-CoA
show the reaction diagram
ATP + suberate + CoA
AMP + diphosphate + suberyl-CoA
show the reaction diagram
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-
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?
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
ATP + 6-carboxyhexanoate + CoA
AMP + diphosphate + 6-carboxyhexanoyl-CoA
show the reaction diagram
ATP + heptanedioate + CoA
?
show the reaction diagram
ATP + heptanedioate + CoA
AMP + diphosphate + heptanedioyl-CoA
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
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32% of the activation relative to Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
2,2'-dipyridyl
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iodoacetic acid
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p-chloromercuribenzoate
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0107 - 0.0705
6-Carboxyhexanoate
0.2996 - 1.5
ATP
0.033 - 0.55
CoA
0.145 - 0.49
heptanedioate
0.529
Heptanoate
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mutant Y211F, pH and temperature not specified in the publication
0.17
MgATP2-
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0.4794
Octanoate
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mutant Y211F, pH and temperature not specified in the publication
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48 - 7.45
6-Carboxyhexanoate
0.0066
adipate
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pH and temperature not specified in the publication
0.44
ATP
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pH and temperature not specified in the publication
0.0001
azelate
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pH and temperature not specified in the publication
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0.87
CoA
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pH and temperature not specified in the publication
0.0051
Glutarate
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pH and temperature not specified in the publication
0.012
Heptanoate
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mutant Y211F, pH and temperature not specified in the publication
0.008
Octanoate
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mutant Y211F, pH and temperature not specified in the publication
0.0038
suberate
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pH and temperature not specified in the publication
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additional information
additional information
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
77.3
freshly prepared enzyme, pH 8.0, at 30ºC
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 9.5
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8: about 55% of maximal activity, 9.5: about 65% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 52
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15°C: about 50% of maximal activity, 52°C: about 15% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3
pH-gradient 6.0-4.0, chromatofocusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28000
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2 * 28000, SDS-PAGE
53000
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x * 53000, SDS-PAGE
60000
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gel filtration
74643
4 * 74643, calculation from sequence of cDNA
298600
calculated from sequence of cDNA
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 53000, SDS-PAGE
homodimer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure determination and analysis of free wild-type AaBioW, selenomethinonine-labeled AaBioW with pimelate, AaBioW with AMP-CPP-Mg2+-pimelate, and AaBioW with CoA-AMP
crystal structure determination and analysis of wild-type enzyme
purified K2PtCl4-pimeloyl-adenylate-diphosphate complex, pimelic acid-CoASH complex, pimeloyl-adenylate-diphosphate, and AMP-PNP-pimelic acid complexes, X-ray diffraction structure determination and analysis at 2.04-2.44 A resolution
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
propen­sity of the purified recombinant enzyme to precipitate and by the loss of activity upon prolonged incubation
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
anion exchange, gel filtration, chromatofocusing
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography, tag cleavage through thrombin, and gel filtration
recombinant His6-tagged enzyme by nickel affinity chromatography, removal of the tag by TEV protease, another step of nickel affinity chromatography, and gel filtration, to homogeneity
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene bioW, encoded in the bio gene cluster, recombinant expression in and complementation of Escherichia coli DELTAbioC DELTA bioH mutant strain STL25, a derivative of strain MG1655
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gene bioW, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta2 (DE3)
gene bioW, sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli
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overexpression in Escherichia coli
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H16A
site-directed mutagenesis, the mutant variant displays only a modest 20% loss in activity relative to the wild-type, reflecting the importance of these other interacting residues in stabilizing CoA binding
R159A
site-directed mutagenesis, the activity to hydrolyze adenylates of noncognate substrates is abolished in the mutant. The R159A variant can no longer proofread, but the enzyme still retains ligase activity and can catalyze the formation of pimeloyl-CoA, the mutant demonstrates a notable reduction in turnover, which is in line with the function of the residue in forming the exterior wall of the pimelate-binding cavity
R201A
site-directed mutagenesis, the mutation has little effect on product formation
R215A
site-directed mutagenesis, the mutant demonstrates a substantial reduction in product formation
S182A
site-directed mutagenesis, the mutant demonstrates a substantial reduction in product formation
Y187A
site-directed mutagenesis, the mutant demonstrates a notable reduction in turnover, which is in line with the function of the residue in forming the exterior wall of the pimelate-binding cavity
Y199A
site-directed mutagenesis, the mutation has little effect on product formation
R213A
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site-directed mutagenesis, almost inactive mutant
R227E
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site-directed mutagenesis, the mutant shows a turnover with the natural substrate pimelic acid that is reduced by around 25fold to about 4% activity remaining compared to the wild-type
R227K
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site-directed mutagenesis, the mutant shows a turnover with the natural substrate pimelic acid that is reduced by around 25fold to about 4% activity remaining compared to the wild-type
Y199F
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site-directed mutagenesis, the mutant retains 55% activity compared to wild-type, the Y199F mutant is inactive with heptanoic acid and octanoic acid, the Y199F mutant displayed a twofold greater activity with pimelic acid but no improvement with azelaic acid compared to wild-type
Y211F
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site-directed mutagenesis, almost inactive mutant; site-directed mutagenesis, the mutant displays activity with both monocarboxylic acid substrates, heptanoic acid and octanoic acid, the Y211F mutant displays about 4fold increased activity with the suberic acid substrate and 3fold increased activity with the azelaic acid substrate relative to the wild-type BioW. The mutant enzymes is also active with 7-bromoheptanoic acid, 7-aminoheptanoic acid, 6-methylheptanoic acid, 7-phenyl­heptanoic acid, and 7-octenoic acid, but not with 7-aminoheptanoic acid; site-directed mutagenesis, the mutant retains 36% activity compared to wild-type
R213A
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site-directed mutagenesis, almost inactive mutant
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Y199F
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site-directed mutagenesis, the mutant retains 55% activity compared to wild-type, the Y199F mutant is inactive with heptanoic acid and octanoic acid, the Y199F mutant displayed a twofold greater activity with pimelic acid but no improvement with azelaic acid compared to wild-type
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Y211F
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site-directed mutagenesis, almost inactive mutant; site-directed mutagenesis, the mutant displays activity with both monocarboxylic acid substrates, heptanoic acid and octanoic acid, the Y211F mutant displays about 4fold increased activity with the suberic acid substrate and 3fold increased activity with the azelaic acid substrate relative to the wild-type BioW. The mutant enzymes is also active with 7-bromoheptanoic acid, 7-aminoheptanoic acid, 6-methylheptanoic acid, 7-phenyl­heptanoic acid, and 7-octenoic acid, but not with 7-aminoheptanoic acid; site-directed mutagenesis, the mutant retains 36% activity compared to wild-type
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additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (402 entries)
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