Information on EC 3.5.1.36 - N-methyl-2-oxoglutaramate hydrolase

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The expected taxonomic range for this enzyme is: Pseudomonas sp.

EC NUMBER
COMMENTARY
3.5.1.36
-
RECOMMENDED NAME
GeneOntology No.
N-methyl-2-oxoglutaramate hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
N-methyl-2-oxoglutaramate + H2O = 2-oxoglutarate + methylamine
show the reaction diagram
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-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
acyl transfer
-
-
with delta-substituted alpha-ketoglutarates, presumably through the formation of an alpha-ketoglutaryl enzyme intermediate
-
hydrolysis of amide bond
-
-
-
-
hydrolysis of ester
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
N-methyl-2-oxoglutaramate methylamidohydrolase
In the reverse reaction, the product cyclizes non-enzymically to 2-hydroxy-N-methyl-5-oxoproline.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5-hydroxy-N-methylpyroglutamate synthase
-
-
-
-
5-hydroxy-N-methylpyroglutamate synthetase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9073-53-4
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain M.A. ATCC No. 23819
-
-
Manually annotated by BRENDA team
Pseudomonas sp. M.A.
strain M.A. ATCC No. 23819
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-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-oxopentanedioic acid 5-benzyl ester + H2O
2-oxoglutarate + benzyl alcohol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
2-oxopentanedioic acid 5-ethyl ester + H2O
2-oxoglutarate + ethanol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
2-oxopentanedioic acid 5-propyl ester + H2O
2-oxoglutarate + propanol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
alpha-ketoglutaramate + H2O
alpha-ketoglutarate + NH3
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
alpha-ketoglutarate + methylamine
N-methyl-2-oxoglutaramate + H2O
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-oxopentanedioic acid 5-benzyl ester + H2O
2-oxoglutarate + benzyl alcohol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
2-oxopentanedioic acid 5-ethyl ester + H2O
2-oxoglutarate + ethanol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
2-oxopentanedioic acid 5-propyl ester + H2O
2-oxoglutarate + propanol
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
alpha-ketoglutaramate + H2O
alpha-ketoglutarate + NH3
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
-
-
-
?
alpha-ketoglutarate + methylamine
N-methyl-2-oxoglutaramate + H2O
show the reaction diagram
Pseudomonas sp., Pseudomonas sp. M.A.
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-
-
?
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-hydroxy-4-methoxy-benzophenone
-
with 1 micromol 2-hydroxy-4-methoxy-benzophenone the activity is 0% after 10 min
Aliphatic alcohols
-
with respect to amines in acyl transfer reactions
butanol
-
competitive inhibitors with respect to methylamine and ethanolamine
DTNB
-
with 20 micromol DTNB the activity is 5% after 10 min
iodoacetamide
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with 20 micromol iodoacetamide the activity is 20% after 10 min
N-Methylmaleimide
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with 20 micromol N-methylmaleimide the activity is 2% after 10 min
NEM
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with 20 micromol NEM the activity is 0% after 10 min
oxalacetate
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a competitive inhibitor
p-hydroxymercurisulfonate
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with 1 micromol p-hydroxymercurisulfonate the activity is 0% after 10 min
Pentanol
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competitive to ethanolamine
Propanol
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competitive to ethanoamine
Tris-HCl
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-
iodoacetate
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with 20 micromol iodoacetate the activity is 33% after 10 min
additional information
-
not inhibited by triethanolamine, 2-methylimidazole, diphosphate and Tricine buffer
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acetone
-
noncompetitive activator with respect to ethanolamine
Aliphatic alcohols
-
7fold increase of hydrolysis of alpha-ketoglutaramate and ethyl alpha-ketoglutarate
Dimethyl formamide
-
noncompetitive activator with respect to ethanolamine
Dioxane
-
noncompetitive activator, effect on the reaction of methylamine with ethyl alpha-ketoglutarate
Organic solvents
-
increase both hydrolysis and transfer reactions involving amines, accelerated up to 7fold
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
alpha-ketoglutaramate
-
-
38
alpha-ketoglutarate
-
-
0.0048
benzyl alpha-ketoglutarate
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-
0.0045
ethyl alpha-ketoglutarate
-
-
46
methylamine
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-
0.0026
propyl alpha-ketoglutarate
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-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0178
alpha-ketoglutarate
-
turnover number based on the molecular weight of 90000
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.98
-
after partial purification
11
-
after purification
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
enzymatic reaction exhibits a pH maximum of about 8
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
90000
-
gel filtration
209087
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
after heated 12 min to 50°C the activity of enzyme is 12%. Untreated enzyme stable
209088
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 20 mM potassium phosphate buffer, pH 7.4, containing 5 mM alpha-ketoglutarate, six months. In the absence of alpha-ketoglutarate the enzyme slowly loses activity over a period of several weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
extraction, addition of streptomycin sulfate, adjust the pH value, centrifugation, ammonium sulfate precipitation, dialysis, purification with DEAE, Sephadex and hydroxyapatite column with dialysis and ultrafiltration steps
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