Information on EC 4.2.1.3 - aconitate hydratase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea

EC NUMBER
COMMENTARY hide
4.2.1.3
-
RECOMMENDED NAME
GeneOntology No.
aconitate hydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Citrate = cis-aconitate + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Carbon fixation pathways in prokaryotes
-
-
Citrate cycle (TCA cycle)
-
-
citric acid cycle
-
-
Glyoxylate and dicarboxylate metabolism
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
citrate(isocitrate) hydro-lyase (cis-aconitate-forming)
Besides interconverting citrate and cis-aconitate, it also interconverts cis-aconitate with isocitrate and, hence, interconverts citrate and isocitrate. The equilibrium mixture is 91% citrate, 6% isocitrate and 3% aconitate. cis-Aconitate is used to designate the isomer (Z)-prop-1-ene-1,2,3-tricarboxylate. An iron-sulfur protein, containing a [4Fe-4S] cluster to which the substrate binds.
CAS REGISTRY NUMBER
COMMENTARY hide
9024-25-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain DPN7T (DSM 17166T, LMG 22922T)
TrEMBL
Manually annotated by BRENDA team
strain DPN7T (DSM 17166T, LMG 22922T)
TrEMBL
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
isoform aconitase A
-
-
Manually annotated by BRENDA team
T
-
-
Manually annotated by BRENDA team
gene citB
-
-
Manually annotated by BRENDA team
strain TM4000
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain D-248
-
-
Manually annotated by BRENDA team
Cucurbita sp.
3 isoforms: Aco I, Aco II and Aco III
-
-
Manually annotated by BRENDA team
strain JRG2387 and JRG3171, contains two major aconitases, AcnA and AcnB
-
-
Manually annotated by BRENDA team
strain W3350
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
bifunctional enzyme with aconitase activity and working as iron-responsive protein. Since iron is required for aconitase activity, but inhibits the RNA-binding activity, the two activities are mutually exclusive
-
-
Manually annotated by BRENDA team
isolate Pb01, gene Pbaco
-
-
Manually annotated by BRENDA team
isolate Pb01, gene Pbaco
-
-
Manually annotated by BRENDA team
Rattus norvegicus Male Fischer 344
strain Male Fischer 344
UniProt
Manually annotated by BRENDA team
Rattus norvegicus Sprague-Dawley
strain Sprague-Dawley
-
-
Manually annotated by BRENDA team
Rheum sp.
i.e. rhubarb
-
-
Manually annotated by BRENDA team
strain MMY011, gene aco1
-
-
Manually annotated by BRENDA team
Saccharomycopsis lipolytica
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
IFO 12371
-
-
Manually annotated by BRENDA team
Streptomyces viridochromogenes strain Tu494
strain Tu494
-
-
Manually annotated by BRENDA team
IFO 4923
-
-
Manually annotated by BRENDA team
IFO 13303
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2R,3S)-2-methylisocitrate
(Z)-2-methylaconitate + H2O
show the reaction diagram
(Z)-2-methylaconitate + H2O
(2R,3S)-2-methylisocitrate
show the reaction diagram
alpha-methyl-cis-aconitate
alpha-methylisocitrate
show the reaction diagram
cis-aconitate
citrate
show the reaction diagram
cis-aconitate
isocitrate
show the reaction diagram
cis-aconitate + H2O
?
show the reaction diagram
cis-aconitate + H2O
citrate
show the reaction diagram
cis-aconitate + H2O
isocitrate
show the reaction diagram
citrate
cis-aconitate
show the reaction diagram
citrate
cis-aconitate + H2O
show the reaction diagram
citrate
isocitrate
show the reaction diagram
isocitrate
?
show the reaction diagram
isocitrate
cis-aconitate
show the reaction diagram
isocitrate
cis-aconitate + H2O
show the reaction diagram
isocitrate
citrate
show the reaction diagram
threo-D-alpha-methylisocitrate
?
show the reaction diagram
Saccharomycopsis lipolytica
-
-
-
-
?
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
(Z)-2-methylaconitate + H2O
(2R,3S)-2-methylisocitrate
show the reaction diagram
cis-aconitate + H2O
isocitrate
show the reaction diagram
citrate
cis-aconitate + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
thiamine diphosphate
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn
-
an X-ray fluorescence measurement performed on a gold-derivative crystal shows the unexpected presence of zinc, in addition to gold and iron
[4Fe-4S] center
-
acon harbors a single unligated iron atom in its [4Fe-4S], enzyme is in this respect unique in mitochondria
additional information
IRP1 is a cytosolic isozyme devoid of labile Fe2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
1,2,3,4-tetracarboxycyclopentane
-
competitive
1,2,3-tricarboxycyclopentene-1
-
competitive
1,3,5-tricarboxypentane
-
competitive
2,2'-dipyridyl
-
noncompetitive
4-hydroxy-2-oxoglutarate
-
competitive
adipate
Saccharomycopsis lipolytica
-
-
ADP
-
inhibition at levels well above its physiological concentration
alpha-picolinic acid
-
noncompetitive
citramalate
citrate
D-glucose 1-phosphate
-
-
D-glucose 6-phosphate
-
-
deferiprone
-
the loss of aconitase activity observed in cells should be ascribed to the chelation of available iron rather than to a direct effect of the chelator on the iron-sulfur clusters of the enzyme
ethyl picolinate
-
isoenzyme is inhibited, isoenzyme I is less or not sensitive
fluoroacetate
Saccharomycopsis lipolytica
-
-
Fluorocitrate
fructose-6-phosphate
fumarate
-
-
GDP
-
inhibition at levels well above its physiological concentration
glyoxylate
-
-
hydrogen peroxide
-
inhibits enzyme activity in cell-free extracts
indomethacin
-
a non-steroidal anti-inflammatory drug, carbonylation of aconitase and release of iron along with the loss of activity in vivo after indomethacin treatment, activation of mitochondrial death pathway by indomethacin, overview
malate
-
-
Maleate
Saccharomycopsis lipolytica
-
-
Mn2+
-
inhibition of enzyme, resulting in up to 90% increase in intracellular citrate. Mitochondrial isoform is significantly more sensitive to Mn2+ than cytosolic isoform. Inhibition leads to conversion of enzyme to iron regulatory protein IRP 1 and increases the abundance of IRP2, leading to reduced H-ferritin expression, inreased transferrin receptor expression, and increased uptake of transferrin. IRP2 has a dominant role in Mn2+-induced alteration of iron homeostasis over aconitase/IRP1
nitric oxide
-
brief exposure leads to a reversible inhibition competitive with isocitrate. subsequently, an irreversible inactivation is observed
nitrosoglutathione
-
irreversible inactivation both in presence and absence of substrate
oxaloacetate
Oxalomalate
oxalomalic acid
-
inhibition of aconitase activity, leading to inhibition of L-glutamate production, L-cystine uptake, and decrease in glutathione concentration in lens epithelial cells and retinal pigment epithelial cells
oxalosuccinate
p-hydroxymercuribenzoate
-
-
peroxynitrite
Phthalic acid
-
competitive
pyromellitic acid
-
competitive
Quinaldic acid
S(1,1,2,2)-tetrafluoroethyl-L-cysteine
inhibition of renal aconitase activity both in vivo and in vitro is a functional consequence of difluorothioamidyl-L-lysine formation by S(1,1,2,2)-tetrafluoroethyl-L-cysteine
Sodium mersalyl
-
-
succinate
-
-
superoxide anion radical
threo-Ls-isocitrate
-
competitive
-
trans-aconitate
tricarballylate
trimellitic acid
-
competitive
trimesic acid
-
competitive
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cysteine
-
enzyme activity increases 2-4fold in presence of both Fe2+ and cysteine. 0.1-1 mM Fe2+ and 0.05-0.5 mM cysteine
Fe4-S4 cluster
ferric ammonium citrate
-
increases activity and gene expression
Hemin
-
increases activity and gene expression
lipoic acid
-
causes an increase in cardiac enzyme activity at doses of 35 and 70 mg/kg of 1.3 and 2.4fold in the animals
menadione sodium bisulfite
-
causes a modest activation of IRP-1 to bind to iron resonsive elements within 15-30 min. Menadione-induced oxidative stress leads to post-translational inactivation of both genetic and enzymatic functions of IRP-1 by a mechanism that lies beyond the classical Fe-S cluster switch and exerts multiple effects on cellular iron metabolism
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.21
(2R,3S)-2-methylisocitrate
-
0.0035 - 0.2
cis-aconitate
0.12 - 11
citrate
0.012 - 3.33
isocitrate
0.0089 - 0.158
methyl-cis-aconitate
0.032 - 0.268
methylisocitrate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.3 - 5.2
cis-aconitate
5.3
citrate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 8, 25C, Lineweaver-Burk method
1.1
isocitrate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 8, 25C, Lineweaver-Burk method
additional information
additional information
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
26
cis-aconitate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 8, 25C, Lineweaver-Burk method
1230
1
citrate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 8, 25C, Lineweaver-Burk method
131
1.2
isocitrate
Salmonella enterica subsp. enterica serovar Typhimurium
-
pH 8, 25C, Lineweaver-Burk method
279
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.81 - 1.9
D-glucose 1-phosphate
1.14 - 2.35
D-glucose 6-phosphate
0.4 - 0.51
fumarate
0.14 - 0.42
glyoxylate
1.81 - 2.94
malate
0.035
nitric oxide
-
reversible inhibition after brief exposure
1.03 - 2.8
oxaloacetate
0.33 - 0.61
succinate
1.37 - 3.41
trans-aconitate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.63 - 6.88
ADP
1.93 - 3.04
ATP
0.007 - 0.024
Fluorocitrate
3.04 - 6.53
GDP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.011 - 0.06
-
aconitate hydratase in blood serum of control rats and rats subjected to myocardial ischemia and treatment with lipoic acid, overview
0.068 - 0.198
-
aconitate hydratase in myocardium of control rats and rats subjected to myocardial ischemia and treatment with lipoic acid, overview
0.206
-
dehydration of citrate to cis-aconitate, pH 8.0, 30C
0.266
-
dehydration of isocitrate to cis-aconitate, pH 8.0, 30C
0.433
-
reverse reaction, hydration of citrate or isocitrate, pH 8.0, 30C
1.11
-
cytoplasmic isoenzyme
2.126
Cucurbita sp.
-
isoenzyme Aco II
2.214
Cucurbita sp.
-
isoenzyme Aco I
3.4
-
mitochondrial isoenzyme
6.13
-
mitochondrial enzyme
11.1
-
cytosolic enzyme
21.4
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8
-
mitochondrial isoenzyme
7.1
-
mitochondrial isoenzyme
7.2
Rheum sp.
-
phosphate buffer
7.2 - 7.8
Saccharomycopsis lipolytica
-
-
7.3
-
soluble enzyme
7.5 - 8
-
isoenzyme I
7.5
-
isoenzyme II
7.7
-
phosphate buffer
7.8 - 8
-
-
8 - 8.5
-
-
8.1
-
N-ethylmorpholine buffer
8.6
-
veronal acetate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
5.5 - 9.2
Saccharomycopsis lipolytica
-
about 35% of maximal activity at pH 5.5 and pH 9.2
6.4 - 8.2
-
pH 6.4: about 60% of maximal activity, pH 8.2: about 75% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 35
-
isoenzyme I
30
-
isoenzyme II
37
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
-
about 60% of maximal activity at 20C and at 40C, isoenzyme I. 20C: about 40% of maximal activity, 40C: about 20% of maximal activity, isoenzyme II
50 - 90
-
50C: about 40% of maximal activity, 90C: about 45% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.24
theoretical value
6.5 - 7.2
-
by IEF-PAGE, identification of 9 different forms with pI-values from 6.5-7.2
7.52
-
recombinant isozyme 1, isoelectric focusing
7.65
-
recombinant isozyme 2, isoelectric focusing
7.79
-
recombinant isozyme 3, isoelectric focusing
8.1
-
calculated
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
mesencephalic-derived cell line
Manually annotated by BRENDA team
-
activity is significantly higher in bone marrow of rats treated with N-nitro-L-arginine methylester
Manually annotated by BRENDA team
-
from fruit juice cells
Manually annotated by BRENDA team
Cucurbita sp.
-
etiolated
Manually annotated by BRENDA team
-
repeated contractions increase aconitase activity by 50%. Increase is not accompanied by increase in aconitase protein, but is markedly inhibited by cyclosporin A
Manually annotated by BRENDA team
-
gastric carcinoma cell line Okajima
Manually annotated by BRENDA team
-
undifferentiated neonatal cardiomyocyte. In heat-shocked cells aconitase activity is reduced
Manually annotated by BRENDA team
Rheum sp.
-
-
Manually annotated by BRENDA team
-
epithelium
Manually annotated by BRENDA team
mitochondrial aconitase is over-expressed in primary mesencephalic cultures
Manually annotated by BRENDA team
purified human CD34+ progenitors derived from granulocyte-colony stimulating factor mobilized peripheral blood cells of normal donors; purified human CD34+ progenitors derived from granulocyte-colony stimulating factor mobilized peripheral blood cells of normal donors
Manually annotated by BRENDA team
-
malignant and nonmalignant tissues
Manually annotated by BRENDA team
-
muscle exercise does not affect aconitase activity despite increased oxidative stress
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
repeated contractionsdo not alter aconitase activity
Manually annotated by BRENDA team
-
activity is significantly higher in spleen of rats treated with N-nitro-L-arginine methylester
Manually annotated by BRENDA team
-
pulvinus bundle sheath cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
Cucurbita sp.
-
very low activity
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
66000
-
gel filtration
67000
-
gel filtration
68500
-
equilibrium sedimentation
75000 - 80000
-
isoenzyme II, gel filtration
79000
-
ultracentrifugation
80900
-
low speed equilibrium sedimentation
84000
-
mitochondrial enzyme, gel filtration
89000
-
equilibrium sedimentation
95000
-
gel filtration
96000
-
cytosolic enzyme, gel filtration
100000
114000
-
recombinant N-terminally His-tagged aconitase, gel filtration
122000
-
recombinant aconitase with an additional Gly at the C-terminus, gel filtration
145000
-
gel filtration
150000 - 160000
-
isoenzyme I, gel filtration
150000
-
gel filtration
additional information
-
partial amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 41000, mitochondrial enzyme, SDS-PAGE; 2 * 45000, cytosolic enzyme, SDS-PAGE
homodimer
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
proteolytic modification
-
the porcine heart enzyme is synthesized as a precursor containing a mitochondrial targeting sequence of 27 amino acid residues which is cleaved to yield a mature enzyme of 754 amino acids
side-chain modification
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of enzyme-fluorocitrate complex
-
2.4 A structure of AcnB, hanging drop vapor diffusion at 17C
-
aconitase form of bifunctional enzyme, evaluation of model for iron regulatory form of enzyme and its binding to iron-responsive elements
recombinant enzyme, hanging-drop vapour-diffusion method. An X-ray fluorescence measurement performed on a gold-derivative crystal shows the unexpected presence of zinc, in addition to gold and iron
-
1.8 A resolution crystal structure of the S642A:citrate complex of mitochondrial aconitase
computational modeling of aconitase inactivation by superoxide and nitric oxide
-
crystallographic evidence for a three-iron center
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
15 h, 50% inactivation, isoenzyme I. 15 h, about 65% loss of activity, isoenzyme II
30
-
5 h, 50% loss of activity, isoenzyme II. 15 h, 90% loss of activity, isoenzyme I. 15 h, 75% loss of activity, isoenzyme II
45
-
complete inactivation after 20 min without fluorocitrate, stable in presence of fluorocitrate 0.05 mM, 40% loss of activity in presence of 20 mM citrate
50
-
30 min, stable
60
-
10 min, complete inactivation
95
-
half-life: 14 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
fluorocitrate protects from inactivation by O2
-
fluorocitrate stabilizes against heat inactivation
-
freezing and thawing inactivates aconitase in absence of glycerol or sucrose
Cucurbita sp.
-
more than 50% of total aconitase activity is lost when cell-free extracts are prepared under air by a French press, by osmotic rupture of spheroplasts or by sonic treatment
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
fluorocitrate protects from inactivation by O2
-
33804
high oxygen sensitivity
-
651095
rapid inactivation by exposure to air
488175
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 25% w/v glycerol, stable for several weeks
Cucurbita sp.
-
-20C, enzyme frozen after dialysis against 0.05 M Tris-HCl, pH 7.5, complete inactivation after 10 days
-
4C, 24 h, pH 5.5-9.0, stable
-
4C, activity of the highly purified cytosolic enzyme decreases by 15-20% during the first 24 h of storage and then by 5% every 24 h, the mitochondrial enzyme decreases 35-40% during the first 24 h and then by 15% every 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AcnA and AcnB
-
enzyme form AcnA
-
isoenzymes Aco I and Aco II
Cucurbita sp.
-
native and SeMet [3Fe-4S] form of AcnB expressed in Escherichia coli
-
native IRP1 from liver
partial
partially, isolation of mitochondria
-
recombinant AcnB5-4 polypeptide
-
recombinant enzymes from Escherichia coli strain BL21(DE3) by gel filtration
-
recombinant His-tagged AcnA by metal affinity chromatography; recombinant His-tagged AcnB by metal affinity chromatography
recombinant His-tagged aconitase from Escherichia coli strain BL21(DE3) by nickel affinty chromatography and gel filtration, removal of the His tag by factor Xa and further purification, or purification of recombinant aconitase, expressed from plasmid pTYB2-acn in Escherichia coli, using chitin beads, overview
-
recombinant IRP1
-
recombinant protein; recombinant protein
using column chromatography
-
using Ni-NTA chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AcnB domain 5-4 polypeptide is expressed in Escherichia coli as a GST-AcnB5-4 fusion protein
-
cytoplasmic aconitase/iron regulatory protein 1 expression analysis, overview
-
expressed in Escherichia coli
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli as functional enzyme
-
expression in Bacillus subtilis
-
expression in Escherichia coli
expression in HEK 293 cells
expression of N-terminally His-tagged aconitase in Escherichia coli strain BL21(DE3), alternative expression method uses overexpression of aconitase from plasmid pTYB2-acn, resulting in an aconitase containing only an additional glycine residue at the C-terminus
-
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha, and in Bacillus subtilis strains GWH1025 and GWH1026, respectively
-
expression of wild-type enzyme and diverse pount and deletion mutants
-
gene AcnA, overexpression of the His-tagged protein in an ASKA library, i.e. the 'a complete set of Escherichia coli K-12 ORF archive library; gene ACnB, overexpression of the His-tagged protein in an ASKA library
gene acnB, expression of wild-type enzyme and the fusion protein ICDH-AcnB in Escherichia coli strain BL21 (DE3)
-
gene ACO1, overexpression in strain H222, high-level expression of ACO in the ACO1 multicopy integrative transformant results in a shift of the citric acid/isocitric acid product pattern into the direction of isocitric acid
-
gene Ccaco1, DNA and amino acid sequence determination, sequence comparisons and phylogenetic analysis; gene Ccaco2, DNA and amino acid sequence determination, sequence comparisons and phylogenetic analysis; gene Ccaco2, DNA and amino acid sequence determination, sequence comparisons and phylogenetic analysis
gene Pbaco, quantitative real-time RT-PCR expression analysis, recombinant expression in Saccharomyces cerevisiae, ACO is present in the extracellular fluid, cell wall enriched fraction, mitochondria, cytosol and peroxisomes of yeast cells
-
overexpression in Escherichia coli, His-tag; overexpression in Escherichia coli, His-tag
the recombinant yeast mitochondrial aconitase is expressed in Escherichia coli as soluble, biologically active enzyme
-
yeast mitochondrial aconitase is expressed in Escherichia coli by cultivation in a bioreactor at different temperatures. Chaperones GroEL and GroES are co-expressed
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
acetate and ethanol as sole carbon sources and in high iron conditions induce the enzyme
acnA expression is enhanced in an acnB mutant
-
citramalate significantly increases citrate content and reduces the mitochondrial isozyme activity, while slightly inducing its protein level, oxalomalate also slightly upregulates the enzyme
-
expression of acnA is repressed by the presence of acnB
-
promoter fusion experiments show that acnA expression is regulated by the cyclic-AMP receptor protein, the fumarate nitrate reduction regulator, the ferric uptake regulator and the superoxide response protein
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C450S
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mutant is enzymatically inactive, glutamate auxotroph and accumulates citrate. Mutant strain exhibits overexpression of the citB promoter and accumulates high levels of aconitase protein. Mutant strain exhibits increased levels of citrate synthase protein. Mutant enzyme does not bind to the citrate synthase 5' leader RNA in vitro
C517A
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enzymatically inactive mutant enzyme that still binds iron responsive elements
R740E/Q744E/F661L/I809T/V852A
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sixfold increase in specific activity compared to wild-type. Mutant strain is defective in sporulation, affecting the expression of deltaK-dependent genes. Accumulation of transcriptional activator GerE mRNa and protein is delayed in the mutant
R741E
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mutant is designed to be defective in RNA binding. Mutant strain is glutamate prototroph and accumulates citrate. Mutant strain exhibits overexpression of the citB promoter and accumulates high levels of aconitase protein. Mutant strain exhibits increased levels of citrate synthase protein. Mutant enzyme does not bind to the citrate synthase 5' leader RNA in vitro
R741E/Q745E
C459S
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catalytically inactive mutant without its [4Fe-4S]-cluster. Mitochondrial morphological defects as a consequence of acon inactivation depend on its [4Fe-4S] cluster
S677A
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catalytically inactive mutant
S711A
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citrate-to-isocitrate aconitase activity is about 70% of the wild-type activity, isocitrate-to-cis-aconitate activity is about 90% of wild-type activity
S711D
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no citrate-to-isocitrate aconitase activity, isocitrate-to-cis-aconitate activity is identical to wild-type activity