Information on EC 6.2.1.1 - acetate-CoA ligase

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

EC NUMBER
COMMENTARY hide
6.2.1.1
-
RECOMMENDED NAME
GeneOntology No.
acetate-CoA ligase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + acetate + CoA = AMP + diphosphate + acetyl-CoA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acid-thiol ligation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
acetate conversion to acetyl-CoA
-
-
acetate fermentation
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Carbon fixation pathways in prokaryotes
-
-
chitin degradation to ethanol
-
-
cis-genanyl-CoA degradation
-
-
ethanol degradation II
-
-
ethanol degradation IV
-
-
Glycolysis / Gluconeogenesis
-
-
Metabolic pathways
-
-
Methane metabolism
-
-
Microbial metabolism in diverse environments
-
-
oxidative ethanol degradation III
-
-
Propanoate metabolism
-
-
propanol degradation
-
-
Pyruvate metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
acetate:CoA ligase (AMP-forming)
Also acts on propanoate and propenoate.
CAS REGISTRY NUMBER
COMMENTARY hide
9012-31-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ATCC 8554
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain ES114, gene acs
-
-
Manually annotated by BRENDA team
strain ES114, gene acs
-
-
Manually annotated by BRENDA team
Amaranthus sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene acsA
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
; DSM 18386
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
DSM 18386
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Methanothermobacter thermautotrophicum
Methanothermobacter thermautotrophicum Z245
strain Z245
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain U
Uniprot
Manually annotated by BRENDA team
strain S
-
-
Manually annotated by BRENDA team
strain 217
-
-
Manually annotated by BRENDA team
strain 217
-
-
Manually annotated by BRENDA team
strain LK2G12
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
synthetic construct
-
-
-
Manually annotated by BRENDA team
Taxus sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
the metalloprotein acetyl-coenzyme A synthase/carbon monoxide dehydrogenase, ACS/CODH, is a bifunctional metalloenzyme found in anaerobic archaea and bacteria that grow hemoautotrophically on CO or CO2, and is significant for biological carbon fixation and understanding the origin of life
malfunction
metabolism
physiological function
additional information
-
growth arrest is caused by elevated Acs activity, while overproduction of ADP-forming Ac-CoA synthesizing systems, EC 6.2.1.13, do not affect the growth behaviour of acetylation-deficient or acetylation-proficient strains, effects of Acs on growth of different strains, also sirtuin-dependent protein acylation/deacylation system-defective strains, overview. Increased CoA biosynthesis partially alleviates the negative effect caused by high Acs activity, regulation, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ADP + acetate + CoA
AMP + diphosphate + acetyl-CoA
show the reaction diagram
ADP + phosphate + acetyl-CoA
ATP + acetate + CoA
show the reaction diagram
-
-
-
-
?
ATP + 3-bromopropanoate + CoA
AMP + diphosphate + 3-bromopropanoyl-CoA
show the reaction diagram
-
-
-
-
-
ATP + 3-chloropropanoate + CoA
AMP + diphosphate + 3-chloropropanoyl-CoA
show the reaction diagram
-
-
-
-
-
ATP + 3-methylvalerate + CoA
AMP + diphosphate + 3-methylvaleryl-CoA
show the reaction diagram
Methanothermobacter thermautotrophicum
-
mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme
-
-
?
ATP + 4-methylvalerate + CoA
AMP + diphosphate + 4-methylvaleryl-CoA
show the reaction diagram
Methanothermobacter thermautotrophicum
-
mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme
-
-
?
ATP + acetate + CoA
?
show the reaction diagram
ATP + acetate + CoA
AMP + diphosphate + acetyl-CoA
show the reaction diagram
ATP + acetate + seleno-CoA
AMP + diphosphate + acetyl-seleno-CoA
show the reaction diagram
-
-
-
-
-
ATP + acrylate + CoA
AMP + diphosphate + acryloyl-CoA
show the reaction diagram
ATP + butyrate + CoA
AMP + diphosphate + butyryl-CoA
show the reaction diagram
ATP + fluoroacetate + CoA
AMP + diphosphate + fluoroacetyl-CoA
show the reaction diagram
ATP + formate + CoA
AMP + diphosphate + formyl-CoA
show the reaction diagram
-
27% of the activity with acetate
-
-
?
ATP + heptanoate + CoA
AMP + diphosphate + heptanoyl-CoA
show the reaction diagram
ATP + hexanoate + CoA
AMP + diphosphate + hexanoyl-CoA
show the reaction diagram
ATP + isobutyrate + CoA
AMP + diphosphate + isobutyryl-CoA
show the reaction diagram
-
28% of the activity with acetate
-
-
?
ATP + methacrylic acid + CoA
AMP + diphosphate + methacryloyl-CoA
show the reaction diagram
-
-
-
-
-
ATP + octanoate + CoA
AMP + diphosphate + octanoyl-CoA
show the reaction diagram
Methanothermobacter thermautotrophicum
-
mutant enzyme W416G catalyzes the reaction, no activity with wild-type enzyme
-
-
?
ATP + pentanoate + CoA
AMP + diphosphate + pentanoyl-CoA
show the reaction diagram
-
6.7% of the activity relative to acetate
-
-
-
ATP + propanoate + CoA
AMP + diphosphate + propanoyl-CoA
show the reaction diagram
ATP + propionate + CoA
AMP + diphosphate + propionyl-CoA
show the reaction diagram
ATP + tetrapolyphosphate
adenosine 5'-pentaphosphate
show the reaction diagram
-
-
-
-
ATP + tripolyphosphate
adenosine 5'-tetraphosphate
show the reaction diagram
-
-
-
-
ATP + valerate + CoA
AMP + diphosphate + valeryl-CoA
show the reaction diagram
CheY + acetyl-CoA + ATP
acetyl-CheY + CoA + AMP + diphosphate
show the reaction diagram
-
CheY is the the excitatory response regulator in the chemotaxis system of Escherichia coli, acetyl-CoA synthetase-catalyzed transfer of acetyl groups from acetate to CheY and autocatalyzed transfer from AcCoA, mechanism, overview
-
-
?
CTP + acetate + CoA
CMP + diphosphate + acetyl-CoA
show the reaction diagram
-
-
-
-
-
dATP + acetate + CoA
dAMP + diphosphate + acetyl-CoA
show the reaction diagram
GTP + acetate + CoA
GMP + diphosphate + acetyl-CoA
show the reaction diagram
-
-
-
-
-
UTP + acetate + CoA
UMP + diphosphate + acetyl-CoA
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
ATP + acetate + CoA
?
show the reaction diagram
ATP + acetate + CoA
AMP + diphosphate + acetyl-CoA
show the reaction diagram
ATP + propionate + CoA
AMP + diphosphate + propionyl-CoA
show the reaction diagram
CheY + acetyl-CoA + ATP
acetyl-CheY + CoA + AMP + diphosphate
show the reaction diagram
-
CheY is the the excitatory response regulator in the chemotaxis system of Escherichia coli, acetyl-CoA synthetase-catalyzed transfer of acetyl groups from acetate to CheY and autocatalyzed transfer from AcCoA, mechanism, overview
-
-
?
dATP + acetate + CoA
dAMP + diphosphate + acetyl-CoA
show the reaction diagram
Q99NB1
AceCS2 plays a role in the production of energy under ketogenic conditions, such as starvation and diabetes. Acetyl-CoAs produced by AceCS2 are utilized mainly for oxidation
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
two types of divalent metal ion requirement, 1. Mg2+, Mn2+, Fe2+, Co2+ or Ca2+ required for the formation of the enzyme-bound acetyl adenylate, 2. Ni2+, Cd2+, Fe2+ or Cu2+ required for adenylate binding
Cu2+
-
two types of divalent metal ion requirement, 1. Mg2+, Mn2+, Fe2+, Co2+ or Ca2+ required for the formation of the enzyme-bound acetyl adenylate, 2. Ni2+, Cd2+, Fe2+ or Cu2+ required for adenylate binding
KCl
-
optimal activity at 1-1.5 M
Li+
-
activation at 5-8 mM, absolute requirement for certain monovalent cations, inhibition above 10 mM
Ni
-
nickel-containing bimetallic site, the bifunctional enzyme carbon monoxide dehydrogenase/acetyl-coenzyme A synthase
Rb+
-
activates, absolute requirement for certain monovalent cations, no inhibition at high concentrations
Tris
-
activates, absolute requirement for certain monovalent cations, no inhibition at high concentrations
additional information
-
the enzyme uses seven metalloclusters in four reaction steps, overview
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
3'-Dephospho-CoASH analogues with a phosphodiester bond
-
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
acetyl-CoA
-
competitive to CoASH
adenylate
-
-
ADP
-
competitive to ATP
Allicin
bicarbonate
-
-
Butyrate
-
propanoate-CoA formation
CO
-
CO inhibits acetyl-CoA synthesis quite strongly and in a cooperative manner
Dicarbonic acid diethyl ester
-
-
diphosphate
erythrose 4-phosphate
-
-
glyceraldehyde 3-phosphate
-
weak
glycerol
glyoxylate
-
-
long-chain acyl-CoA compounds
Monovalent cations
-
at 200 mM
-
NaCl
-
concentration of 5-20 mM decrease the activity 20-25%
Ni
-
the authors favor a mechanism in which methylation occurs first to Ni(p0 -) or Ni(pI -)[Fe4S4]+, followed by coordination of CO to form Ni(pII)(CO)(CH3) which breaks one of the S(Nid) bonds (forming the bis square planar Ni(II) species, as if the Ni(d)N2S2 unit were acting as a biological pseudodiphosphine, mimicking behavior common to a bidentate phosphine). The CO-insertion/CH3-migration occurs on one metal forming the trigonal planar Ni(pII)-acetyl intermediate. Finally, addition of thiolate produces the thioester. The authors disfavor the unprecedented bimetallic, CO-insertion/CH3-migration mechanism (both in its diamagnetic and paramagnetic guise) and disfavors CO, CH3+, or thiolate (CoA) binding to the distal Ni. Finally, Ni in the proximal site produces a better catalyst than does Cu
Nonidet P40
-
weak
O2
-
the enzyme is O2-sensitive
p-chloromercuribenzoate
-
-
p-hydroxymercuribenzoate
-
inhibition is reversible by either CoA or mercaptoethanol
P1,P5-di(adenosine-5)pentaphosphate
Methanothermobacter thermautotrophicum
-
inhibits ADP formation
palmitoyl-CoA
Propanoate
-
butyryl-CoA formation
pyridoxal 5'-phosphate
-
-
Seleno-CoA
-
competitive to CoA
Short-chain CoA esters
-
-
-
sorbitol
Sucrose
Tween 100
-
-
-
Xylulose 5-phosphate
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
stimulates
acetate
CobB Sir2 protein
-
activation of the acetylated enzyme requires the nicotinamide adenine dinucleotide-dependent protein deacetylase activity of the CobB Sir2 protein
-
DTT
-
stimulates
GSH
-
stimulates
reduced ferredoxin
-
required
SIR2 protein
-
short-chain fatty acid activation by acyl-coenzyme A synthetases requires SIR2 protein function
-
SIRT1
-
AceCS1 is completely inactivated upon acetylation and is rapidly reactivated by SIRT3 deacetylation
-
SIRT3
-
AceCS2 is completely inactivated upon acetylation and is rapidly reactivated by SIRT3 deacetylation
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
29.2
2-methylvalerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
7.6
4-methylvalerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
0.04 - 11000
acetate
0.0037 - 1.8
acetyl-CoA
5.26 - 17
Acrylate
0.093 - 0.4
ADP
0.017 - 17
ATP
0.46 - 151.9
Butyrate
0.011 - 1.2
CoA
31.2
fluoroacetate
-
-
10.2
Heptanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
6.1
hexanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
0.65 - 6.6
MgATP2-
18.1
Octanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
0.23 - 0.272
phosphate
3.1 - 10.5
Propanoate
3.9 - 188.8
Propionate
4.7
tripolyphosphate
-
synthesis of adenosine 5-tetraphosphate
11.1
valerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.67
2-methylvalerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
10.9
4-methylvalerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
0.3 - 9400
acetate
0.7 - 100.6
ATP
0.013 - 10.8
Butyrate
0.3 - 144.9
CoA
7.3
Heptanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
5.9
hexanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
3.3
Octanoate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
5.9 - 46.3
Propionate
11.9
valerate
Methanothermobacter thermautotrophicum
-
65C, mutant enzyme W416G
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.29 - 229
acetate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.7
acetyl-CoA
-
pH 8.0
12
ADP
-
pH 8.0
15
AMP
-
pH 8.0
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
Ni-activated A110C mutant alpha subunit at 1 atm CO
8.1
-
Ni-activated A222L mutant alpha subunit at 1 atm CO
11.9
AceCS2
13.4
-
formation of acetyl-CoA
26.8
AceCS1
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2
-
CO/acetyl-CoA exchange assay at
6.3
-
adenosine tetraphosphate synthesis
7 - 7.5
Methanothermobacter thermautotrophicum
assay at
8.2
Methanothermobacter thermautotrophicum
-
-
8.3 - 10.2
-
in 25 mM KCl buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8.5
-
pH 5.5: about 60% of maximal activity, pH 8.5: about 50% of maximal activity
6 - 10.5
-
6: about 50% of maximal activity, 10.5: about 60% of activity maximum
6 - 9.7
-
6.0: about 45% of maximal activity, 9.7: about 70% of maximal activity
6.5 - 8.5
-
30-40% of maximal activity at pH 6.5 and 8.5
6.8 - 10.1
-
about 50% of maximal activity at pH 6.8 and pH 10.1
6.8 - 8.8
-
about 50% of maximal activity at pH 6.8 and 8.8
7.3 - 8.1
-
90% of maximal activity at pH 7.3 and 8.1
7.5 - 10
-
7.5: about 50% of maximal activity, 10.0: about 70% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27
-
CO/acetyl-CoA exchange assay at
65 - 70
65
Methanothermobacter thermautotrophicum
-
80
-
arsenolytic assay at and temperature optimum
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 50
-
20C: about 45% of maximal activity, 50C: about 30% of maximal activity
45 - 85
-
45C: about 55% of maximal activity, 85C: about 60% of maximal activity, AF-ACS2
45 - 75
Methanothermobacter thermautotrophicum
45C: about 55% of maximal activity, 75C: about 50% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
marked induction of AceCS1 mRNA and protein during differentiation of 3T3-L1 cells, neither AceCS2 mRNA nor protein is detected in undifferentiated or differentiated 3T3-L1 cells
Manually annotated by BRENDA team
-
cells express higher levels of cytosolic acetyl-CoA synthetase ACSS2 under hypoxia than normoxia. Knockdown of ACSS2 by RNA interference in tumor cells enhances tumor cell death under long-term hypoxia in vitro. The ACSS2 suppression slows tumor growth in vivo. Tumor cells excrete acetate and the quantity increases under hypoxia, the pattern of acetate excretion follows the expression pattern of ACSS2. The ACSS2 knockdown leads to a corresponding reduction in the acetate excretion in tumor cells
Manually annotated by BRENDA team
-
from Glycine max L. cv. Williams inoculated with Bradyrhizobium japonicum
Manually annotated by BRENDA team
-
cells express higher levels of cytosolic acetyl-CoA synthetase ACSS2 under hypoxia than normoxia. Knockdown of ACSS2 by RNA interference in tumor cells enhances tumor cell death under long-term hypoxia in vitro. The ACSS2 suppression slows tumor growth in vivo. Tumor cells excrete acetate and the quantity increases under hypoxia, the pattern of acetate excretion follows the expression pattern of ACSS2. The ACSS2 knockdown leads to a corresponding reduction in the acetate excretion in tumor cells
Manually annotated by BRENDA team
-
cells express higher levels of cytosolic acetyl-CoA synthetase ACSS2 under hypoxia than normoxia. Knockdown of ACSS2 by RNA interference in tumor cells enhances tumor cell death under long-term hypoxia in vitro. The ACSS2 suppression slows tumor growth in vivo. Tumor cells excrete acetate and the quantity increases under hypoxia, the pattern of acetate excretion follows the expression pattern of ACSS2. The ACSS2 knockdown leads to a corresponding reduction in the acetate excretion in tumor cells
Manually annotated by BRENDA team
-
cells express higher levels of cytosolic acetyl-CoA synthetase ACSS2 under hypoxia than normoxia. Knockdown of ACSS2 by RNA interference in tumor cells enhances tumor cell death under long-term hypoxia in vitro. The ACSS2 suppression slows tumor growth in vivo. Tumor cells excrete acetate and the quantity increases under hypoxia, the pattern of acetate excretion follows the expression pattern of ACSS2. The ACSS2 knockdown leads to a corresponding reduction in the acetate excretion in tumor cells
Manually annotated by BRENDA team
-
newly-pupated pupae of both sexes
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
AceCS2, high activity
Manually annotated by BRENDA team
AceCS2, high activity
Manually annotated by BRENDA team
additional information
no AceCS2 activity is detected in liver. Marked induction of AceCS1 mRNA and protein during differentiation of 3T3-L1 cells, neither AceCS2 mRNA nor protein is detected in undifferentiated or differentiated 3T3-L1 cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
AceCS1
Manually annotated by BRENDA team
-
isoform ACS2 is localized primarily to the nucleus, with a minor amount in the cytosol
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Cryptococcus neoformans var. grubii serotype A (strain H99 / ATCC 208821 / CBS 10515 / FGSC 9487)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
58000
-
gel filtration
60000
-
gel filtration
72000
-
gel filtration
80000
gel filtration
139000
-
gel filtration
141000
gel filtration
144800
Methanothermobacter thermautotrophicum
gel filtration
148000
-
gel filtration
150000
-
gel filtration
151000
-
sedimentation equilibrium analysis
221200
-
AF-ACS2, gel filtration
250000
-
low speed sedimentation without reaching equilibrium
251200
-
gel filtration
261800
-
ultracentrifugal studies, sedimentation equilibrium absorption optics
610000
-
analytical ultracentrifugation
625000
-
gel filtration
670000
-
; gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterotetramer
-
alpha2beta2
homooligomer
monomer
octamer
-
8 * 75000, SDS-PAGE
tetramer
trimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
proteolytic modification
the putative mitochondrial targeting signal is cleaved during the transportation of the enzyme into the mitochondria matrix
side-chain modification
-
the acetyltransferase enzyme, AcuA, controls the activity of the acetyl coenzyme A synthetase, AcsA, by acetylating residue Lys549, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, 2.3 A resolution, residues 72-713 of ACS (subcloned into expression vector pET28a and expressed in Escherichia coli) in complex with AMP
vapor diffusion method, crystallographic structures of wild-type enzyme and mutant enzymes R194A, R584A, R584E, K609A, and V386A
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
30 min, 15% loss of activity
60
-
5 min, 20% loss of activity
90
-
120 min, stable
98
-
half-life: 72 min
100
-
half-life: 8 min. Addition of 1 M (NH4)2SO4 stabilizes the enzyme to a half-life of 24 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ATP and AMP stabilize
-
ATP and glycerol stabilize during storage at 0-4C
-
denatured upon freezing
-
freezing and thawing causes loss of activity
-
KCl stabilizes
-
more than 90% loss of activity in solutions of low ionic strength, for 20 min, at 0-4C or at room temperature
-
particularly sensitive to repeated freezing
-
photoinactivation in presence of methylene blue
-
storage stability is enhanced by high salt and protein concentration
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme is O2-sensitive
-
687824
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.2 mM DTT, stable for at least one month
-
-20C, 0.5 M potassium phosphate, pH 7.5, 7 mM 2-mercaptoethanol and 0.5 mM EDTA, protein concentration above 5 mg/ml, no loss of activity after 6 months
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-20C, cannot be stored for more than 3 days without a considerable loss of activity
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-20C, stable for several months
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0-4C, 20 mM Tris/Hcl, pH 8.0, 10 mM ATP, 5 mM MgCl2, 1 mM DTT or 5 mM 2-mercaptoethanol, 20% glycerol, stable for at least 1 month
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4C, 0.1 M buffer, pH 7.5, 3 mM mercaptoethanol, stable for several weeks
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4C, 20% w/v glycerol, stable for 1 month
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partial
-
recombinant alpha and beta subunits from Escherichia coli strain BL21(DE3) by heat tteatment at 90C for 30 min, hydrophobic interaction chromatography, and gel filtration
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recombinant C-terminally His-tagged ACS from Escherichia coli strain BL21(DE3) by nickel affinity chromatography under anaerobic conditions
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recombinant enzyme
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
10-deacetylbaccatin III 10beta-O-acetyltransferase (DBAT) from Taxus can catalyze the transfer of acetyl, propionyl or n-butyryl from CoA to the C10-hydroxyl of 10-deacetylbaccatin III. Escherichia coli JM109 are transformed to recombinantly express dbat, and this enzyme function is coupled to that of acetyl-CoA synthase expressed from and regulated by genes encoded on the bacterial chromosome
Taxus sp.
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acetyl-CoA synthetase overexpression in Escherichia coli demonstrates more efficient acetate assimilation and lower acetate accumulation
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expression in Escherichia coli
expression in Escherichia coli, AF-ACS2
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expression of alpha-subunit in Escherichia coli
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expression of C-terminally His-tagged ACS in Escherichia coli strain BL21(DE3)
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genes acdIa and acdIb encoding the alpha and beta subunits, expression in Escherichia coli strain BL21(DE3)
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overexpression in Escherichia coli
production in HEK -293 cells
residues 72-713 of ACS are subcloned into expression vector pET28a and expressed in Escherichia coli
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of CO dehydrogenase/acetyl coenzyme A synthase in Methanosarcina spp. is coordinately regulated in response to substrate by differential transcription initiation and early elongation termination near the 3' end of a 371-bp leader sequence
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A498A
Methanothermobacter thermautotrophicum
alteration does not significantly affect the Km for any substrate but reduces the turnover rate kcat 41fold
A500T
Methanothermobacter thermautotrophicum
kcat value decreases 44fold
D502A
Methanothermobacter thermautotrophicum
inactive mutant enzyme
D502E
Methanothermobacter thermautotrophicum
inactive mutant enzyme
D502N
Methanothermobacter thermautotrophicum
inactive mutant enzyme
G501A
Methanothermobacter thermautotrophicum
two- to threefold reduced Km values for acetate, ATP and CoA. The turnover rate is over 200fold reduced
I312A
Methanothermobacter thermautotrophicum
-
kcat/Km for acetate is 15fold lower than wild-type value, kcat/Km for propionate is 5.2fold lower than wild-type value. Mutant enzyme shows activity with butyrate
T313K
Methanothermobacter thermautotrophicum
-
Km for acetate is 170fold higher than wild-type value, kcat for acetate is 34fold lower than wild-type value
T313V
Methanothermobacter thermautotrophicum
-
kcat/Km for acetate is 2.5fold lower than wild-type value, kcat/Km for propionate is identical to wild-type value
T499A
Methanothermobacter thermautotrophicum
kcat value decreases 83fold
V388A
Methanothermobacter thermautotrophicum
-
kcat/Km for acetate is 6.9fold lower than wild-type value, kcat/Km for propionate is 2.5fold higher than wild-type value, mutant enzyme shows activity with wild-type enzyme
V388G
Methanothermobacter thermautotrophicum
-
kcat/Km for acetate is 93fold lower than wild-type value, kcat/Km for propionate is 26fold lower than wild-type value
W416G
Methanothermobacter thermautotrophicum
-
kcat/Km for acetate is 71.5fold lower than wild-type value, kcat/Km for propionate is 13fold lower than wild-type value, mutant enzyme shows activity with: butyrate, valerate, hexanoate, heptanoate, octanoate, 4-methylvalerate and 3-methylvalerate
Y498F
Methanothermobacter thermautotrophicum
inactive mutant enzyme
I312A
Methanothermobacter thermautotrophicum Z245
-
kcat/Km for acetate is 15fold lower than wild-type value, kcat/Km for propionate is 5.2fold lower than wild-type value. Mutant enzyme shows activity with butyrate
-
T313K
Methanothermobacter thermautotrophicum Z245
-
Km for acetate is 170fold higher than wild-type value, kcat for acetate is 34fold lower than wild-type value
-
T313V
Methanothermobacter thermautotrophicum Z245
-
kcat/Km for acetate is 2.5fold lower than wild-type value, kcat/Km for propionate is identical to wild-type value
-
V388A
Methanothermobacter thermautotrophicum Z245
-
kcat/Km for acetate is 6.9fold lower than wild-type value, kcat/Km for propionate is 2.5fold higher than wild-type value, mutant enzyme shows activity with wild-type enzyme
-
V388G
Methanothermobacter thermautotrophicum Z245
-
kcat/Km for acetate is 93fold lower than wild-type value, kcat/Km for propionate is 26fold lower than wild-type value
-
A110C
-
site-directed mutagenesis, alpha-subunit mutant, which does not show cooperative CO inhibition in contrast to the wild-type enzyme
A222L
-
site-directed mutagenesis, alpha-subunit mutant, which does not show cooperative CO inhibition in contrast to the wild-type enzyme
A265M
-
site-directed mutagenesis, alpha-subunit mutant, the recombinantly expressed mutant enzymes cannot be purified
C509A
-
site-directe mutagenesis, mutant C509A shows a significantly diminished methyl transfer activity compared to the wild-type enzyme
C509H
-
site-directed mutagenesis, mutant C509H can accept a methyl group from CH3-Co3+FeSP at over 70% extent. The near-wild-type-level of methyl group transfer activity for C509H indicates that the di-nickel site is assembled well in this mutant, and strongly suggests that an imidazole group can bridge the di-nickel site to the cubane of the A-cluster. Histidine that replaces the bridging cysteine 509 might function as a bridge, with one nitrogen of the imidazole ring coordinating to a cubane Fe and the other nitrogen coordinating to Nip
C509S
-
site-directed mutagenesis, mutant C509S, in which the cysteinate bridge C509 might be replaced by a serine oxide, exhibits no detectable methyl transfer activity. Oxygen is a harder donor than sulfide, and the electronic coupling between the cubane and the di-nickel site may differ relative to sulfide. Absence of methyl transfer activity in C509S indicates that an O bridge is not sufficient for this communication
C509V
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site-directed mutagenesis, mutant C509V exhibits no detectable methyl group transfer activity due to it lacking a bridging coordinating atom, Val is more bulky and has greater steric hindrance
D212Ebeta
-
site-directed mutagenesis, the mutant shows 2-4% of wild-type activity, slightly impaired in arsenolysis
D212Nbeta
-
site-directed mutagenesis, the mutant shows highly reduced phosphorylation/phosphorolysis activity, but only slightly impaired in arsenolysis
E218Qalpha
-
site-directed mutagenesis, inactive mutant
H257Dalpha
-
site-directed mutagenesis, inactive mutant
H71Abeta
-
site-directed mutagenesis, inactive mutant concerning phosphorylation/phosphorolysis, slightly impaired in arsenolysis
A357V
-
kcat/Km for ATP is 1.2fold higher than wild-type value, kcat/Km for CoA is 3.2fold lower than wild-type value
D517G
-
kcat/Km for ATP is 6.5fold lower than wild-type value, kcat/Km for CoA is 9.5fold lower than wild-type value
D517P
-
kcat/Km for ATP is 1.4fold lower than wild-type value, kcat/Km for CoA is 23.7fold lower than wild-type value
G266S
-
random mutagenesis, mutant Km for acetate is 268fold higher than that of the AcsWT enzyme, while kcat is 3fold reduced; the Acs mutant does not cause growth arrest in contrast to the wild-type enzyme
G524L
-
inactive mutant enzyme; mutant enzyme is unable to catalyze the complete reaction yet catalyzes the adenylation half-reaction with activity comparable to the wild-type enzyme
G524S
-
kcat/Km for ATP is 1.6fold lower than wild-type value, kcat/Km for CoA is 19fold lower than wild-type value
R194A
-
kcat/Km for ATP is 1.1fold higher than wild-type value, kcat/Km for CoA is 6.3fold lower than wild-type value
R194E
-
kcat/Km for ATP is 1.2fold lower than wild-type value, kcat/Km for CoA is 4.75fold lower than wild-type value
R526A
-
kcat/Km for ATP is 1.2fold higher than wild-type value, kcat/Km for CoA is 9.5fold lower than wild-type value
R584A
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kcat/Km for ATP is 1.2 fold than wild-type value, kcat/Km for CoA is 19fold lower than wild-type value
R584E
-
kcat/Km for ATP is 1.1fold higher than wild-type value, kcat/Km for CoA is 21fold lower than wild-type value
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
colorimetric assay method to measure acetyl-CoA synthetase activity
medicine
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