Information on EC 5.4.99.13 - isobutyryl-CoA mutase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
5.4.99.13
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RECOMMENDED NAME
GeneOntology No.
isobutyryl-CoA mutase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-methylpropanoyl-CoA = butanoyl-CoA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
intramolecular transfer reaction
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SYSTEMATIC NAME
IUBMB Comments
2-methylpropanoyl-CoA CoA-carbonylmutase
This bacterial enzyme utilizes 5'-deoxyadenosylcobalamin as a cofactor. Following substrate binding, the enzyme catalyses the homolytic cleavage of the cobalt-carbon bond of AdoCbl, yielding cob(II)alamin and a 5'-deoxyadenosyl radical, which initiates the the carbon skeleton rearrangement reaction by hydrogen atom abstraction from the substrate. At the end of each catalytic cycle the 5'-deoxyadenosyl radical and cob(II)alamin recombine, regenerating the resting form of the cofactor. The enzyme is prone to inactivation resulting from occassional loss of the 5'-deoxyadenosyl molecule. Inactivated enzymes are repaired by the action of EC 2.5.1.17, cob(I)yrinic acid a,c-diamide adenosyltransferase, and a G-protein chaperone, which restore cob(II)alamin (which is first reduced to cob(I)alamin by an unidentified reductase) to 5'-deoxyadenosylcobalamin and load it back on the mutase. Some mutases are fused with their G-protein chaperone. These enzyme can also catalyse the interconversion of isovaleryl-CoA with pivalyl-CoA.
CAS REGISTRY NUMBER
COMMENTARY hide
116405-23-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
isolated from methyl t-butyl ether-contaminated groundwater, able to grow on methyl t-butyl ether and ethyl t-butyl ether as sole carbon sources
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Manually annotated by BRENDA team
isolated from methyl t-butyl ether-contaminated groundwater, able to grow on methyl t-butyl ether and ethyl t-butyl ether as sole carbon sources
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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
2-hydroxyisobutyrate
3-hydroxybutyrate
show the reaction diagram
2-methylpropanoyl-CoA
butanoyl-CoA
show the reaction diagram
isobutyryl-CoA
n-butyryl-CoA
show the reaction diagram
isobutyrylcarba(dethia)-CoA
butyrylcarba(dethia)-CoA
show the reaction diagram
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
2-methylpropanoyl-CoA
butanoyl-CoA
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
coenzyme A
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
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or cobalamin, strictly required. During growth on limiting concentrations of cobalt, strain accumulates 2-hydroxyisobutyrate
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
GDP
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the presence of GDP decreases kcat about 1.6-1.7fold while increasing the Km about 2fold. Consequently, the catalytic efficiency kcat/Km value of IcmF decreases 3fold in the presence of GDP
GTP
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the presence of GTP decreases kcat about 1.6-1.7fold while increasing the Km about 2fold. Consequently, the catalytic efficiency kcat/Km value of IcmF decreases 4.5fold in the presence of GTP
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.054
2-methylpropanoyl-CoA
pH 7.4, 30C
0.057
Butanoyl-CoA
pH 7.4, 30C
0.0201 - 0.0508
isobutyryl-CoA
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
38.2
Butanoyl-CoA
pH 7.4, 30C
1.88 - 3.1
isobutyryl-CoA
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
37 - 150
isobutyryl-CoA
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.023
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with butanoyl-CoA as the substrate from extracts of Streptomyces cinnamonensis
0.34
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recombinant enzyme, at 37C
0.75
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recombinant enzyme, at 37C
1
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activity reconstituted by mixing recombinant His6-IcmA produced in Escherichia coli with IcmB isolated from Streptomyces cinnamonensis
1.1
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recombinant enzyme, at 37C
3.3
the specific activity varies with the molar ratio of IcmA to IcmB
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Cupriavidus metallidurans (strain ATCC 43123 / DSM 2839 / NBRC 102507 / CH34)
Cupriavidus metallidurans (strain ATCC 43123 / DSM 2839 / NBRC 102507 / CH34)
Cupriavidus metallidurans (strain ATCC 43123 / DSM 2839 / NBRC 102507 / CH34)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
14300
alpha2beta2, IcmA2IcmB2, 2 * 62500 + 2 * 14300, gel filtration, IcmB provides the cobalamin-binding domain
62500
alpha2beta2, IcmA2IcmB2, 2 * 62500 + 2 * 14300, gel filtration, IcmB provides the cobalamin-binding domain
65000
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IcmA, 1 * 65000 + several proteins of lower mass, SDS-PAGE
153000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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IcmA, 1 * 65000 + several proteins of lower mass, SDS-PAGE
tetramer
alpha2beta2, IcmA2IcmB2, 2 * 62500 + 2 * 14300, gel filtration, IcmB provides the cobalamin-binding domain
additional information
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IcmB contains coenzyme B12 binding domain
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
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the enzyme is unstable at 60C
additional information
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stable at room temperature for several hours
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA-Sepharose column chromatography, Source 15Q column chromatography, and Superdex 200 gel filtration
partial, using ammonium sulfate precipitation, and DEAE-Sepharose column chromatography
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recombinant IcmB is purified using Q-Sepharose, and two Mono Q column chromatography
using ammonium sulfate precipitation, DEAE, Q-sepharose, superdex, gel electrophoresis, and B12 affinity column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme small subunit
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expressed in Escherichia coli BL21(DE3) cells
IcmA, expression in Escherichia coli and Streptomyces lividans, IcmA shows 70% of homology to the methylmalonyl-CoA mutase large subunit from Streptomyces cinnamonensis
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IcmB, expression in Escherichia coli, IcmB shows significant identity to several cobalamin-dependent proteins
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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disruption of IcmA in Streptomyces cinnamonensis
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