Information on EC 2.8.3.12 - glutaconate CoA-transferase

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The expected taxonomic range for this enzyme is: Firmicutes

EC NUMBER
COMMENTARY
2.8.3.12
-
RECOMMENDED NAME
GeneOntology No.
glutaconate CoA-transferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
show the reaction diagram
ping-pong mechanism
-
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
show the reaction diagram
ping-pong mechanism; reaction mechanism
-
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
show the reaction diagram
reaction mechanism
-
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
coenzyme A transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Butanoate metabolism
-
glutamate degradation V (via hydroxyglutarate)
-
Microbial metabolism in diverse environments
-
Styrene degradation
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-CoA:(E)-glutaconate CoA-transferase
Glutarate, (R)-2-hydroxyglutarate, propenoate and propanoate, but not (Z)-glutaconate, can also act as acceptors.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
(E)-glutaconate CoA-transferase
-
-
-
-
coenzyme A-transferase, glutaconate
-
-
-
-
GCT
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
79078-99-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
5-part of the hydroxyglutarate operon with gctAB genes encoding Gct, ATCC 25085
Q59111 and Q59112
UniProt
Manually annotated by BRENDA team
Clostridium symbiosum HB 25
HB 25
-
-
Manually annotated by BRENDA team
no activity in Clostridium tetanomorphum
-
-
-
Manually annotated by BRENDA team
no activity in Clostridium tetanomorphum H1
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
-
r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
Q59111 and Q59112
-
-
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
-
only 1-isomer is formed
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
very good substrate, broad substrate specificity
-
?, r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
removal of 3'-phospho group from acetyl-CoA leads to reduced activity
-
?, r
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
involved in glutamate fermentation via hydroxyglutarate pathway, activates glutaconate to glutaconyl-CoA prior to glutaconate decarboxylation
-
?
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
Clostridium symbiosum HB 25
-
-
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
-
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
-
-
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
-
-
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
Q59111 and Q59112
-
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
-
-
in vitro formation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA, the 1-isomer is kinetically favoured, the 5-isomer is thermodynamically favoured
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-1-CoA
show the reaction diagram
-
-
in vitro formation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA, the 1-isomer is kinetically favoured, the 5-isomer is thermodynamically favoured, product in vivo, 5-isomer only occurs in vitro, (R)-2-hydroxyglutaryl-1-CoA is an excellent substrate in the reverse reaction
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
Q59111 and Q59112
involved in the conversion of (R)-2-hydroxyglutarate to crotonyl-CoA in the pathway of glutamate fermentation
-
?
acetyl-CoA + acrylate
acrylyl-CoA + acetate
show the reaction diagram
-
poor substrate
-
-
?
acetyl-CoA + crotonate
crotonyl-CoA + acetate
show the reaction diagram
-
poor substrate
-
-
?
acetyl-CoA + isocrotonate
isocrotonyl-CoA + acetate
show the reaction diagram
-
poor substrate
-
-
?
glutaconyl-CoA + (R)-2-hydroxyglutarate
glutaconate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
-
formation of the 5-isomer, not of the 1-isomer
?
glutaconyl-CoA + (R)-2-hydroxyglutarate
glutaconate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
very good substrate
generation of both isomers: (R)-2-hydroxyglutaryl-1-CoA and (R)-2-hydroxyglutaryl-5-CoA
?
glutaconyl-CoA + (S)-2-hydroxyglutarate
glutaconate + (S)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
at 60% of the rate with (R)-isomer or glutarate
-
-
?
glutaconyl-CoA + 3-hydroxyglutarate
glutaconate + 3-hydroxyglutaryl-CoA
show the reaction diagram
-
at 20% of the rate with glutarate
-
-
?
glutaconyl-CoA + adipate
glutaconate + adipyl-CoA
show the reaction diagram
-
at 60% of the rate with glutarate
-
-
?
glutaconyl-CoA + butyrate
butyryl-CoA + glutaconate
show the reaction diagram
-
-
-
-
?
glutaconyl-CoA + glutarate
glutaconate + glutaryl-CoA
show the reaction diagram
-
very good substrate
-
?
glutaconyl-CoA + propionate
propanoyl-CoA + glutaconate
show the reaction diagram
-
moderate substrate
-
-
?
glutaryl-CoA + acetate
glutarate + acetyl-CoA
show the reaction diagram
-
-
-
?
glutaryl-CoA + acetate
glutarate + acetyl-CoA
show the reaction diagram
-
-
-
r
glutaryl-CoA + acetate
glutarate + acetyl-CoA
show the reaction diagram
Q59111 and Q59112
-
-
?
glutaryl-CoA + acetate
glutarate + acetyl-CoA
show the reaction diagram
-
exchange of oxygen atoms between the substrates and the catalytic residue betaE-54, exchange is site-specific, reversible and requires both substrates, catalytic mechanism
-
r
additional information
?
-
-
no substrate: lactone-CoA
-
-
-
additional information
?
-
-
wild-type enzyme has significant hydrolase activity using acetyl-CoA as substrate, but no hydrolysis of glutaryl-CoA
-
-
-
additional information
?
-
-
residue E-54 of the subunit GctB is directly involved in catalysis by formation of a CoASH ester intermediate
-
-
-
additional information
?
-
Q59111 and Q59112
residue E-54 of the small subunit GctB is directly involved in catalysis
-
-
-
additional information
?
-
-
no substrates: (Z)-glutaconate, acetyl-4'-phosphopantetheine, C4-dicarboxylic acids, reverse reaction not with glutamate, 2-oxoglutarate, succinate, malate or citrate
-
-
-
additional information
?
-
-
GCT structure, the active site is located at the interface of subunits A and B and is formed by loops of both subunits
-
-
-
additional information
?
-
-
catalytic residue is E-54 of subunit B, catalytic mechanism
-
-
-
additional information
?
-
-
not involved in the dehydration of (R)-2-hydroxyglutarate to glutaconate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl-CoA + (E)-glutaconate
acetate + glutaconyl-1-CoA
show the reaction diagram
-
involved in glutamate fermentation via hydroxyglutarate pathway, activates glutaconate to glutaconyl-CoA prior to glutaconate decarboxylation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
?
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
-
in the course of glutamate fermentation
-
r
acetyl-CoA + (R)-2-hydroxyglutarate
acetate + (R)-2-hydroxyglutaryl-CoA
show the reaction diagram
Q59111 and Q59112
involved in the conversion of (R)-2-hydroxyglutarate to crotonyl-CoA in the pathway of glutamate fermentation
-
?
additional information
?
-
-
not involved in the dehydration of (R)-2-hydroxyglutarate to glutaconate
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NaBH4
-
10 mM, in presence of 0.4 mM glutyral-CoA, at 25C, 30 min, almost complete inhibition
4-hydroxymercuribenzoate
-
0.2 mM, at 37C, 2 min, 80% loss of activity, 2-mercaptoethanol partially restores activity
additional information
-
not inhibited by 2 mM iodoacetate, 2 mM iodoacetamide or 1 mM 5,5-dithiobis(2-nitrobenzoate)
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.5
-
(R)-2-hydroxyglutarate
-
cosubstrate glutaconyl-CoA
0.035
-
(R)-2-hydroxyglutaryl-1-CoA
-
pH 7, 37C, cosubstrate acetate
14
-
(S)-2-Hydroxyglutarate
-
cosubstrate glutaconyl-CoA
13
-
3-hydroxyglutarate
-
cosubstrate glutaconyl-CoA
26
-
acetate
-
cosubstrate glutaconyl-CoA
0.17
-
acetyl-CoA
-
cosubstrate glutaconate
10
-
Acrylate
-
cosubstrate acetyl-CoA
8
-
Adipate
-
cosubstrate glutaconyl-CoA
150
-
Butyrate
-
cosubstrate glutaconyl-CoA
500
-
crotonate
-
cosubstrate acetyl-CoA
0.2
-
glutaconate
-
pH 7, 25C, cosubstrate acetyl-CoA
0.017
-
glutaconyl-CoA
-
cosubstrate glutarate
0.7
-
Glutarate
-
cosubstrate glutaconyl-CoA
0.015
-
glutaryl-CoA
-
pH 7, 25C, cosubstrate acetate
0.028
-
glutaryl-CoA
-
pH 7, cosubstrate acetate, wild-type enzyme
0.064
-
glutaryl-CoA
-
pH 7, cosubstrate acetate, 65 kDa GctF fusion protein
100
-
isocrotonate
-
cosubstrate acetyl-CoA
16
-
Propionate
-
cosubstrate glutaconyl-CoA
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
39
-
glutaryl-CoA
-
pH 7, cosubstrate acetate, 65 kDa GctF fusion protein
140
-
glutaryl-CoA
-
pH 7, cosubstrate acetate, wild-type enzyme
additional information
-
additional information
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
Q59111 and Q59112
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
-
assay at; optimum, glutaconyl-CoA + acetate
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.6
7.4
-
about 70% of maximal activity at pH 5.6 and 7.4
8.5
-
-
about 10% of maximal activity at pH 8.5
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
assay at, hydrolase activity
37
-
-
assay at, CoA-transferase activity
PDB
SCOP
CATH
ORGANISM
Acidaminococcus fermentans (strain ATCC 25085 / DSM 20731 / VR4)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
260000
-
-
heterooctamer GCT
275000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
octamer
-
alpha4beta4, 4 * 32000 + 4 * 34000, SDS-PAGE
octamer
Q59111 and Q59112
alpha4beta4, 4 * 35725 + 4 * 29168, GctA and GctB, calculated from the DNA sequences; alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
octamer
-
alpha4beta4, 4 * 36000 + 4 * 29000, GctA and GctB, SDS-PAGE
octamer
-
alpha4beta4, 4 * 35000 + 4 * 29000, structure and association of the subunits A and B
octamer
-
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry
octamer
-
(alphabeta)4
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
betaE54D mutant enzyme, expressed in Escherichia coli
-
native and recombinant enzyme, expressed in Escherichia coli
Q59111 and Q59112
recombinant GCT, expressed in Escherichia coli, sitting-drop vapour diffusion method
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, crystalline suspension in ammonium sulfate, 2 years, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
betaE54D mutant enzyme and 65 kDa GctF fusion protein, expressed in Escherichia coli
-
native and recombinant enzyme, expressed in Escherichia coli
Q59111 and Q59112
recombinant GCT, expressed in Escherichia coli
-
recombinant wild-type and betaE54D mutant enzyme, expressed in Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
GCT expression in Escherichia coli
-
gctAB genes encoding the two subunits GctA and GctB are cloned, sequenced and overexpressed together in Escherichia coli DH5alpha, the two genes are located at the beginning of the hydroxyglutarate operon, GctA: 320 amino acids protein, GctB: 266 amino acids protein
Q59111 and Q59112
overexpression of betaE54N and betaE54A mutant enzyme in Escherichia coli, fusion of the genes gctA and gctB, encoding the 2 subunits of Gct, yields GctF, which is expressed in Escherichia coli as a 65 kDa protein with 30% of wild-type activity; overexpression of wild-type and betaE54D mutant enzyme in Escherichia coli
-
overexpression of wild-type and betaE54D mutant enzyme in Escherichia coli
-
wild-type and mutant genes gctA and gctB encoding the two subunits GctA and GctB are cloned and expressed together in Escherichia coli XL1-Blue
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E54A
-
0.02% as active as wild-type enzyme; mutation in the subunit GctB, inactive mutant
E54A
-
mutation in the subunit GctB, inactive mutant
E54A
-
mutant without CoA-transferase or acyl-CoA hydrolase activity; mutation in the subunit GctB, inactive mutant
E54D
-
above 7% as active as wild-type enzyme, mutant with changed catalytic mechanism without CoASH intermediate, the Escherichia coli strain producing mutant enzyme shows a lower growth rate and reduced amount of recombinant enzyme; mutation in the subunit GctB
E54D
-
mutation in the subunit GctB; replacement of the catalytic glutamate by aspartate converts the mutant enzyme to a thiol ester hydrolase
E54D
-
mutation decreases CoA-transferase and increases acyl-CoA hydrolase activity, mechanism, mutant catalyses the hydrolysis of glutaryl-CoA, acetyl-CoA and 3-butenoyl-CoA, mutant Escherichia coli strain shows reduced growth rate after induction; mutation in the subunit GctB
E54N
-
mutation in the subunit GctB, inactive mutant
E54N
-
mutant without CoA-transferase or acyl-CoA hydrolase activity; mutation in the subunit GctB, inactive mutant
E54Q
-
mutant activity increases from 1% to almost 100% upon incubation with acetyl-CoA and glutaconate at 37C within 40 h, the substrates induce the conversion of the mutant glutamine residue into the glutamate residue of the wild-type enzyme; mutation in the subunit GctB
E54Q
-
1% as active as wild-type enzyme, by incubating with both substrates for 20 h at room temperature, glutamine is completely converted to glutamate yielding a fully active CoA-transferase; mutation in the subunit GctB
E64A
-
mutation in the subunit GctB, 30% as active as wild-type enzyme