Information on EC 1.3.1.85 - crotonyl-CoA carboxylase/reductase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.3.1.85
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RECOMMENDED NAME
GeneOntology No.
crotonyl-CoA carboxylase/reductase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(2S)-ethylmalonyl-CoA + NADP+ = (E)-but-2-enoyl-CoA + CO2 + NADPH + H+
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylation
reductive carboxylation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
crotonyl-CoA/ethylmalonyl-CoA/hydroxybutyryl-CoA cycle (engineered)
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ethylmalonyl-CoA pathway
Glyoxylate and dicarboxylate metabolism
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
(2S)-ethylmalonyl-CoA:NADP+ oxidoreductase (decarboxylating)
The reaction is catalysed in the reverse direction. This enzyme, isolated from the bacterium Rhodobacter sphaeroides, catalyses (E)-but-2-enoyl-CoA-dependent oxidation of NADPH in the presence of CO2. When CO2 is absent, the enzyme catalyses the reduction of (E)-but-2-enoyl-CoA to butanoyl-CoA, but with only 10% of maximal activity (relative to (E)-but-2-enoyl-CoA carboxylation).
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
crotonyl-CoA carboxylase/reductase, a key enzyme of the ethylmalonyl-CoA pathway for acetyl-CoA assimilation, is not essential for growth with 3-hydroxypropionate
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2E)-3-cyclohexylprop-2-enoyl-CoA + NADPH + CO2 + H+
(2S)-2-carboxy-3-cyclohexylpropanoyl-CoA + NADP+
show the reaction diagram
(2E)-4-methylhex-2-enoyl-CoA + NADPH + CO2 + H+
[(2R)-2-methylbutyl]propanedioyl-CoA + NADP+
show the reaction diagram
(2E)-5-chloropent-2-enoyl-CoA + NADPH + CO2 + H+
(2S)-2-carboxy-5-chloropentanoyl-CoA + NADP+
show the reaction diagram
(2E)-5-methylhex-2-enoyl-CoA + NADPH + CO2 + H+
(2S)-(3-methylbutyl)propanedioyl-CoA + NADP+
show the reaction diagram
(2E)-6-methyloct-2-enoyl-CoA + NADPH + CO2 + H+
(2S)-2-carboxy-6-methyloctanoyl-CoA + NADP+
show the reaction diagram
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?
(2E)-hept-2-en-6-ynoyl-CoA + NADPH + CO2 + H+
(2S)-2-craboxy-hept-6-ynoyl-CoA + NADP+
show the reaction diagram
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?
(2E)-hex-2-enoyl-CoA + NADPH + CO2 + H+
(S)-butylproanedioyl-CoA + NADP+
show the reaction diagram
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?
(2E)-oct-2-enoyl-CoA + NADPH + CO2 + H+
(2S)-2-carboxyoctanoyl-CoA + NADP+
show the reaction diagram
(E)-crotonyl-CoA + CO2 + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
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100% relative specific activity, the carboxylation reaction is the physiologically relevant reaction
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r
(E)-crotonyl-CoA + NADPH + H+
butyryl-CoA + NADP+
show the reaction diagram
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10% of maximal activity (compared with vmax of crotonyl-CoA carboxylation)
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?
3-indolylacryloyl-CoA + NADPH + CO2 + H+
indolylmethylmalonyl-CoA + NADP+
show the reaction diagram
4-coumaroyl-CoA + NADPH + CO2 + H+
(2S)2-carboxy-3-(4-hydroxyphenyl)propionyl-CoA + NADP+
show the reaction diagram
acryloyl-CoA + CO2 + NADPH + H+
methylmalonyl-CoA + NADP+
show the reaction diagram
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acryloyl-CoA is accepted as an alternative substrate analogue by the enzyme with 40% relative activity (compared with vmax of crotonyl-CoA carboxylation)
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?
cinnamoyl-CoA + NADPH + CO2 + H+
(2S)2-carboxy-3-phenylpropionyl-CoA + NADP+
show the reaction diagram
crotonyl-CoA + CO2 + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
crotonyl-CoA + CO2 + NADPH + H+
?
show the reaction diagram
crotonyl-CoA + NADPH + CO2 + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
crotonyl-CoA + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
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?
crotonyl-CoA + NADPH + H+
?
show the reaction diagram
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?
crotonyl-CoA + NADPH + H+
butyryl-CoA + NADP+
show the reaction diagram
decanoyl-CoA + NADPH + H+
?
show the reaction diagram
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?
hexanoyl-CoA + NADPH + H+
?
show the reaction diagram
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?
octanoyl-CoA + NADPH + H+
?
show the reaction diagram
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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
(E)-crotonyl-CoA + CO2 + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
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100% relative specific activity, the carboxylation reaction is the physiologically relevant reaction
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r
crotonyl-CoA + CO2 + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
crotonyl-CoA + CO2 + NADPH + H+
?
show the reaction diagram
crotonyl-CoA + NADPH + H+
(2S)-ethylmalonyl-CoA + NADP+
show the reaction diagram
Q6T2B7
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetate
activity is upregulated during photoheterotrophic growth on acetate compared to the activities during growth on succinate
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2
(2S)-ethylmalonyl-CoA
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in 100 mM Tris-HCl buffer (pH 7.9), temperature not specified in the publication
0.2 - 0.4
(E)-crotonyl-CoA
0.14 - 0.2
4-Coumaroyl-CoA
0.5
acryloyl-CoA
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pH and temperature not specified in the publication
0.089 - 0.38
cinnamoyl-CoA
0.2
CO2
0.4 - 1.58
crotonyl-CoA
0.7
NADPH
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
104
(2S)-ethylmalonyl-CoA
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in 100 mM Tris-HCl buffer (pH 7.9), temperature not specified in the publication
0.0735 - 0.109
4-Coumaroyl-CoA
0.0688 - 0.0877
cinnamoyl-CoA
0.0675 - 0.1755
crotonyl-CoA
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
520
(2S)-ethylmalonyl-CoA
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in 100 mM Tris-HCl buffer (pH 7.9), temperature not specified in the publication
0.525 - 0.545
4-Coumaroyl-CoA
0.18 - 0.985
cinnamoyl-CoA
0.0583 - 0.257
crotonyl-CoA
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47000
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2 * 47000, gel filtration
47700
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2 * 47700, gel filtration
105000
143000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, using 100 mM CAPS-NaOH pH 10.0, 38% (v/v) PEG 400
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE-Sepharose column chromatography and Cibacron Blue 3GA agarose column chromatography
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Ni-NTA column chromatography and Superdex 200HR gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BLR(DE3) cells
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expressed in Escherichia coli JM101 cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
although the reaction is reversible, the activity of crotonyl-CoA carboxylase/reductase is up-regulated at least 60fold for Rhodobacter sphaeroides during growth with acetate versus succinate
TetR-type activator CcrR regulates the expression of crotonyl-CoA reductase/carboxylase. A ccrR null mutant strain ismpaired in its ability to grow on C1 and C2 compounds, correlating with the reduced activity of crotonyl-CoA reductase/carboxylase. The activity of the promoter required for ccr expression decreases as much as 50% in the absence of ccrR compared to wild-type. CcrR directly binds to the region upstream of the katA-ccr promoter
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A182G
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inactive
A182L
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the mutant with increased activity selectively catalyzes carboxylation over the regular reduction reaction
V350A
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the mutant shows increased activity compared to the wild type enzyme
V350F
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the mutant shows increased activity compared to the wild type enzyme
V350G
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the mutant shows increased activity compared to the wild type enzyme
A182G
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inactive
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A182L
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the mutant with increased activity selectively catalyzes carboxylation over the regular reduction reaction
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V350A
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the mutant shows increased activity compared to the wild type enzyme
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V350F
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the mutant shows increased activity compared to the wild type enzyme
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V350G
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the mutant shows increased activity compared to the wild type enzyme
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