Information on EC 3.5.1.3 - omega-amidase

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

EC NUMBER
COMMENTARY hide
3.5.1.3
-
RECOMMENDED NAME
GeneOntology No.
omega-amidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a monoamide of a dicarboxylate + H2O = a dicarboxylate + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
transamidation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Alanine, aspartate and glutamate metabolism
-
-
aspartate and asparagine metabolism
-
-
glutamate and glutamine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
omega-amidodicarboxylate amidohydrolase
Acts on glutaramate, succinamate and their 2-oxo derivatives.
CAS REGISTRY NUMBER
COMMENTARY hide
9025-19-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene At5g12040
UniProt
Manually annotated by BRENDA team
gene At5g12040
UniProt
Manually annotated by BRENDA team
168, wild-type
-
-
Manually annotated by BRENDA team
168, wild-type
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
chicken
-
-
Manually annotated by BRENDA team
lettuce
-
-
Manually annotated by BRENDA team
wild-type strain, 74-A
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
spinach
-
-
Manually annotated by BRENDA team
gram-negative extreme thermophilic bacterium; YT-1
-
-
Manually annotated by BRENDA team
YT-1
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
2-hydroxysuccinamate + H2O
malate + NH3
show the reaction diagram
2-oxoglutaramate + H2O
2-oxoglutarate + NH3
show the reaction diagram
2-oxoglutaramate + hydroxylamine
2-oxoglutaryl hydroxamate + NH3
show the reaction diagram
2-oxoglutaramate + methylamine
?
show the reaction diagram
2-oxosuccinamate + H2O
2-oxosuccinate + NH3
show the reaction diagram
2-oxosuccinamate + H2O
oxaloacetate + NH3
show the reaction diagram
asparagine + H2O
aspartate + NH3
show the reaction diagram
-
-
-
-
?
butyl glutarate + hydoxylamine
glutaramate + butanol
show the reaction diagram
-
hydroxaminolysis
-
-
?
caproamide + H2O
caproic acid + NH3
show the reaction diagram
-
no hydroxamte formation
-
-
?
delta-ethyl 2-oxoglutarate + H2O
?
show the reaction diagram
delta-methyl 2-oxoglutarate + H2O
?
show the reaction diagram
ethyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
ethyl glutarate + hydoxylamine
glutaramate + ethanol
show the reaction diagram
-
hydroxaminolysis
-
-
?
ethyl glutarate + methylamine
?
show the reaction diagram
ethyl succinate + H2O
?
show the reaction diagram
-
-
-
-
?
ethyl succinate + hydoxylamine
succinamate + ethanol
show the reaction diagram
-
hydroxaminolysis
-
-
?
ethyl succinate + methylamine
?
show the reaction diagram
fumarate + H2O
?
show the reaction diagram
-
-
-
-
?
fumarate + hydroxylamine
fumaryl hydroxamate + H2O
show the reaction diagram
-
acyl transfer reaction
-
-
?
gamma-ethyl 2-oxoglutarate + H2O
?
show the reaction diagram
gamma-ethyl 2-oxoglutarate + methylamine
?
show the reaction diagram
gamma-methyl 2-oxoglutarate + H2O
?
show the reaction diagram
gamma-methyl 2-oxoglutarate + methylamine
?
show the reaction diagram
gamma-monomethyl-2-oxoglutaramate + H2O
gamma-monomethyl-2-oxoglutarate + NH3
show the reaction diagram
-
tested at pH 7.2 and pH 8.5
-
-
?
glutaramate + H2O
glutarate + NH3
show the reaction diagram
glutaramate + hydroxylamine
glutaryl hydroxamate + NH3
show the reaction diagram
glutaramate + methanol
glutarate methyl ester + hydroxylamine
show the reaction diagram
-
methanolysis
-
-
?
glutaramate + methylamine
?
show the reaction diagram
glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
glutarate + hydroxylamine
glutaryl hydroxamate
show the reaction diagram
L-asparagine + hydroxylamine
L-asparaginyl hydroxamate + H2O
show the reaction diagram
-
amide transfer reaction
-
-
?
L-isoasparagine + H2O
L-aspartate + NH3
show the reaction diagram
-
poor substrate
-
-
?
L-isoglutamine + H2O
L-glutamate + NH3
show the reaction diagram
-
poor substrate
-
-
?
L-leucine amide + H2O
L-Leu + NH3
show the reaction diagram
-
no hydroxamte formation
-
-
?
L-phenylalanine amide + H2O
L-Phe + NH3
show the reaction diagram
-
poor substrate, 12% compared with alpha-ketoglutaramate
-
-
?
malate + H2O
?
show the reaction diagram
malate + hydroxylamine
malyl hydroxamate + H2O
show the reaction diagram
methyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
methyl glutarate + hydoxylamine
glutaramate + methanol
show the reaction diagram
methyl glutarate + methylamine
?
show the reaction diagram
methyl succinate + H2O
?
show the reaction diagram
-
-
-
-
?
methyl succinate + hydoxylamine
succinamate + methanol
show the reaction diagram
-
hydroxaminolysis
-
-
?
methyl succinate + methylamine
?
show the reaction diagram
N-methyl-2-oxoglutaramate + H2O
2-oxoglutarate + methylamine
show the reaction diagram
p-chlorophenyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
p-methoxyphenyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
p-methylphenyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
p-methylphenyl glutarate + hydroxylamine
glutaramate + 4-methylphenol
show the reaction diagram
-
-
-
-
?
p-methylphenyl glutarate + methylamine
?
show the reaction diagram
-
-
-
-
?
phenyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
propyl glutarate + H2O
?
show the reaction diagram
-
-
-
-
?
propyl glutarate + hydoxylamine
glutaramate + propanol
show the reaction diagram
-
hydroxaminolysis
-
-
?
succinamate + H2O
succinate + NH3
show the reaction diagram
-
tested at pH 7.2 and pH 8.5
-
-
?
succinamate + H2O
succinic acid + NH3
show the reaction diagram
succinamate + hydroxylamine
succinyl hydroxamate + NH3
show the reaction diagram
succinamate + methylamine
?
show the reaction diagram
-
transamidation
-
-
?
succinamide + H2O
succinate + NH3
show the reaction diagram
succinamide + hydroxylamine
succinyl hydroxamate + ?
show the reaction diagram
succinate + H2O
?
show the reaction diagram
-
-
-
-
?
succinate + hydroxylamine
succinyl hydroxamate + H2O
show the reaction diagram
succinimide + hydroxylamine
?
show the reaction diagram
succinyl hydroxamate + H2O
succinate + hydroxylamine
show the reaction diagram
succinylmonohydroxamate + H2O
succinate + hydroxylamine
show the reaction diagram
-
-
-
-
r
valeramide + H2O
valeric acid + NH3
show the reaction diagram
-
poor substrate
-
-
?
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
2-oxoglutaramate + H2O
2-oxoglutarate + NH3
show the reaction diagram
2-oxosuccinamate + H2O
2-oxosuccinate + NH3
show the reaction diagram
-
alpha-ketosuccinamate
-
-
?
2-oxosuccinamate + H2O
oxaloacetate + NH3
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,2,2-Trifluoroethanol
-
more than 2 M: irreversible inactivation
2-Chloroethanol
-
more than 2 M: irreversible inactivation
5,5'-dithiobis-(2-nitrobenzoic acid)
6-diazo-5-oxo-L-norleucine
-
DON, completely
alpha-ketosuccinamate
-
10 mM: 75% inhibition
dioxane
-
up to 5% dioxane: no effect on hydrolysis of ethyl alpha-keto-glutarate or glutaramate, at 10% dioxane: slight, 5-10% inhibition
ethanol
Glutaramate
-
competitive inhibition, 10 mM glutaramate, hydrolysis of methyl alpha-ketoglutarate: 43% inhibition, hydrolysis of ethyl alpha-ketoglutarate: 34% inhibition
glycylglycine
-
competitive inhibitor, Ki: 5 mM
iodoacetamide
iodoacetate
methylamine
N-ethylmaleimide
p-mercuribenzoate
phosphate
succinamate
-
competitive inhibition, 5 mM succinamate, hydrolysis of methyl alpha-ketoglutarate: 57% inhibition, hydrolysis of ethyl alpha-ketoglutarate: 42% inhibition
sulfhydryl reagents
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
hydroxylamine
-
uncompetitive activation
methanol
-
uncompetitive activation
Sodium deoxycholate
-
membrane-associated form
sodium lauryl sulfate
-
membrane-associated form
Triton X-100
-
membrane-associated form
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.58
2-hydroxysuccinamate
pH 8.5, 30C, recombinant His-tagged enzyme
-
0.0054 - 9
2-Oxoglutaramate
3.3
2-oxoglutarate
-
hydrolysis
0.003 - 6.13
2-oxosuccinamate
9.9 - 31
ethyl glutarate
8.3 - 17
ethyl succinate
0.06
gamma-ethyl 2-oxoglutarate
-
hydrolysis
0.03
gamma-methyl 2-oxoglutarate
-
hydrolysis
0.012
gamma-monomethyl-2-oxoglutaramate
-
pH 7.2, 30C
1.27 - 25
Glutaramate
1 - 100
hydroxylamine
5000
methanol
-
with substrate, acyl donor: glutaramate
1.8 - 52
methyl glutarate
0.7 - 8
methyl succinate
30 - 55
methylamine
3.7
p-chlorophenyl glutarate
-
hydrolysis
8
p-methoxyphenyl glutarate
-
hydrolysis
2
p-methylphenyl glutarate
-
hydrolysis
8
phenyl glutarate
-
hydrolysis
6.8 - 18
propyl glutarate
0.14 - 17
succinamate
30 - 63
succinate
0.8
succinyl hydroxamate
-
hydrolysis
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
17.5
2-Oxoglutaramate
Mus musculus
-
pH 8.5, 30C
0.87 - 1.15
2-oxosuccinamate
134.5
gamma-monomethyl-2-oxoglutaramate
Mus musculus
-
pH 7.2, 30C
4.1 - 8.8
Glutaramate
2 - 2.8
succinamate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
89.7
2-Oxoglutaramate
Mus musculus
-
pH 8.5, 30C
881
51.2 - 385
2-oxosuccinamate
2391
11200
gamma-monomethyl-2-oxoglutaramate
Mus musculus
-
pH 7.2, 30C
41355
2.8 - 6.9
Glutaramate
3342
8.7 - 20
succinamate
3493
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
17.4
-
ethyl alpha-ketoglutarate hydrolysis
17.67
-
liver, mitochondrial enzyme
42.4
-
liver, soluble, cytoplasmic, enzyme
126
-
2-oxoglutaramate as substrate, pH 8.5, 37C
570
-
2-oxosuccinamate as substrate, pH 8.5, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
hydroxamate formation, substrate glutarate or succinate
6.5
-
acyl transferase activity, sharp optimum
6.5 - 7.5
-
broad pH-optimum, substrate glutaramate or succinamate; hydroxaminolysis
6.5 - 9.5
-
broad pH-optimum, hydrolysis of alpha-ketosuccinamate
6.5
-
acyl transferase activity, sharp optimum
7 - 9
-
amide transferase activity, broad optimum
7 - 9
-
amide transferase activity, broad optimum
7.2
-
very low activity on 2-oxoglutaramte as substrate compared to assay at pH 8.5, presumably because Nit2 only acts on the linear forms of this substrate. The generation of the linear form from the predominant, cyclic form is pH-dependent, being much fatsre at pH 8.5 than at pH 7.2
7.4
-
ethyl alpha-keto-glutarate hydrolysis, assay at
8.5 - 9
-
pH-optimum for hydroxaminolysis of alpha-ketoglutaramate
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45 - 56
-
broad temperature optimum
70
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 63
-
about 50% of maximum activity
63 - 87
-
about 50% of maximum activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
-
cerebral omega-amidase activity is relatively high at embryonic day 5 but lower between days 5 and 17, at embryonic day 23 activity rises to a maximum; embryo brain
Manually annotated by BRENDA team
-
cardiac muscle and skeletal muscle
Manually annotated by BRENDA team
-
S1-S3 region of nephron
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
-
1 * 31000 + 1 * 26000, or 2 * 31000, not distinguishable, calculated from sequence, the 26000 Da subunit is a glutathione S-transferase subunit, the 31000 Da the catalytic subunit of omega-amidase
27000
-
1 * 30000 + 1 * 27000, or 2 * 30000, not distinguishable, SDS-PAGE, the 27000 Da subunit is a glutathione S-transferase subunit, the 30000 Da the catalytic subunit of omega-amidase
28000
-
liver, cytoplasmic enzyme, 2 * 28000
34000
x * 40300, about, sequence calculation, x * 34000, recombinant His-tagged enzyme without transit peptide, SDS-PAGe
36000 - 38500
-
gel filtration
40300
x * 40300, about, sequence calculation, x * 34000, recombinant His-tagged enzyme without transit peptide, SDS-PAGe
60000
-
gel filtration on Sephadex G-200
62000
-
gel filtration
120000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
2 * 30000, SDS-PAGE, there is no glutathione S-transferase isoform like determined for the Nit2 enzyme in rat
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
20 min, 90% loss of activity
288892
5
-
20 min, 32% loss of activity
288892
6 - 10
-
20 min, 30C, stable between, no irreversible inactivation
288892
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
20-30% glycerol or 2-mercaptoethanol stabilize
-
does not require mercaptoethanol or succinate for stability
enzyme quite stable
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2,2,2-trifluoroethanol
-
more than 2 M: unstable to
2-chloroethanol
-
more than 2 M: unstable to
Ethanol
-
more than 2 M: unstable to
Methanol
-
more than 2 M: stable to and uncompetitive activation
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
soluble enzyme, sensitive to oxidation, full activity easily regenerated by addition of 5 mM 2-mercaptoethanol
-
288884
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, protein concentration above 0.1 mg/ml, 0.05 M potassium phosphate, pH 7.2, 0.05 M KCl, 3 months, quite stable
-
-20C, protein concentration above 0.1 mg/ml, 3 months, quite stable
-
3C, 1 mM dithiothreitol, 2 weeks, less than 10% activity lost
-
cytoplasmic and mitochondrial enzyme, 20% glycerol, 5 mM 2-mercaptoethanol, pH 7.2
-
enzyme with high specific activity, 30% glycerol, 0.8 mg protein/ml, 11 years, still active
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, dialysis, DE-52 chromatography, hydroxylapatite chromatography, purified 3600fold, 15.1% recovery
-
from rat liver
-
liver, cytoplasm
-
liver, cytosolic and mitochondrial enzyme
-
partial, membrane-associated, with detergents
-
recombinant N-terminally His-tagged enzyme lacking the chloroplast transit peptide from Escherichia coli by nickel affinity chromatography and desalting gel filtration
recombinant protein, purified to homogeneity on a HisTrap column
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as a fusion ptotein with an N-terminal polyHis tag in Escherichia coli and purified to homogeneity on a HisTrap column
-
gene At5g12040, DNA and amino acid sequence determination and analysis, expression of N-terminally His-tagged recombinant enzyme lacking the chloroplast transit peptide, commencing at Met63, in Escherichia coli
gene nit, the gene is encoded in the nic-gene cluster located on plasmid pAO1 and flanked by mobile elements
-
Nit2-amplified DNA is obtained by PCR of Nit2 cDNA clnoed from human liver cDNA
-
the gene is encoded in the nic-gene cluster on the chromosome
-
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme sensitive to oxidation, full activity easily regenerated in a few min by addition of 5 mM 2-mercaptoethanol
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
-
development of a method to synthesize 2-oxoglutaramate which is not commercially available and development of an enzyme assay method to measure the hydrolysis of 2-oxoglutaramate and 2-oxosuccinamate in a 96-well plate format
medicine
-
in patients with liver disease and encephalopathy there is a good correlation between degree of neurological dysfunction and increase in alpha-ketoglutaramate in cerebrospinal fluid, omega-amidase is suitable for determination of alpha-ketoglutaramate