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

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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 ACCESSION NO.
LITERATURE
N-methyl-2-oxoglutaramate + H2O = 2-oxoglutarate + methylamine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
-
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
5-hydroxy-N-methylpyroglutamate synthase
-
-
-
-
5-hydroxy-N-methylpyroglutamate synthetase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9073-53-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain M.A. ATCC No. 23819
-
-
Manually annotated by BRENDA team
Pseudomonas sp. M.A.
strain M.A. ATCC No. 23819
-
-
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
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 ACCESSION NO.
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.
-
-
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
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
-
with 20 micromol iodoacetamide the activity is 20% after 10 min
N-Methylmaleimide
-
with 20 micromol N-methylmaleimide the activity is 2% after 10 min
NEM
-
with 20 micromol NEM the activity is 0% after 10 min
oxalacetate
-
a competitive inhibitor
p-hydroxymercurisulfonate
-
with 1 micromol p-hydroxymercurisulfonate the activity is 0% after 10 min
Pentanol
-
competitive to ethanolamine
Propanol
-
competitive to ethanoamine
iodoacetate
-
with 20 micromol iodoacetate the activity is 33% after 10 min
additional information
-
not inhibited by triethanolamine, 2-methylimidazole, diphosphate and Tricine buffer
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
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
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3
-
alpha-ketoglutaramate
-
-
38
-
alpha-ketoglutarate
-
-
0.0048
-
benzyl alpha-ketoglutarate
-
-
0.0045
-
ethyl alpha-ketoglutarate
-
-
46
-
Methylamine
-
-
0.0026
-
propyl alpha-ketoglutarate
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0178
-
alpha-ketoglutarate
-
turnover number based on the molecular weight of 90000
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.98
-
-
after partial purification
11
-
-
after purification
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
-
enzymatic reaction exhibits a pH maximum of about 8
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
90000
-
-
gel filtration
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
after heated 12 min to 50°C the activity of enzyme is 12%. Untreated enzyme stable
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
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
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-