Information on EC 3.4.11.B5 - Thermococcus onnurineus deblocking aminopeptidase

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The expected taxonomic range for this enzyme is: Thermococcus onnurineus

EC NUMBER
COMMENTARY hide
3.4.11.B5
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
Thermococcus onnurineus deblocking aminopeptidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Catalytic efficiency for hydrolysis of N-leucyl 4-nitroanilide is 105fold higher than catalytic efficiency for hydrolysis of N-acetyl-L-leucyl 4-nitroanilide. Catalytic efficiency for hydrolysis of L-methionyl 4-nitroanilide is similar to 5fold higher compared to catalytic efficiency for hydrolysis of l-leucyl 4-nitroanilide
show the reaction diagram
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SYSTEMATIC NAME
IUBMB Comments
A7J1X4 {UniProt}
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-leucyl 4-nitroanilide + H2O
L-leucine + 4-nitroaniline
show the reaction diagram
L-methionyl 4-nitroanilide + H2O
L-methionine + 4-nitroaniline
show the reaction diagram
N-acetyl-L-leucyl 4-nitroanilide + H2O
N-acetyl-L-leucine + 4-nitroaniline
show the reaction diagram
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catalytic efficiency for hydrolysis of Leu-4-nitroanilide is 105fold higher than catalytic efficiency for hydrolysis of acetyl-Leu-4-nitroanilide
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
activates; activates, relative activity is about 90% compared to Mn2+
Cu2+
relative activity is about 30% compared to Mn2+; relative activity is about 35% compared to Mg2+
Mg2+
activates; activates, relative activity is about 80% compared to Mn2+
Mn2+
activates; relative activity is about 75% compared to Mg2+
NaCl
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stimulates, optimal cobcentration
Ni2+
relative activity is about 40% compared to Mn2+; relative activity is about 80% compared to Mg2+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
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0.5 mM, complete inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.8 - 37.5
L-leucyl 4-nitroanilide
0.2 - 10.9
L-methionyl 4-nitroanilide
1.1
N-acetyl-L-leucyl 4-nitroanilide
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pH 7.5, 80°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
37.5 - 108
L-leucyl 4-nitroanilide
10.9 - 101
L-methionyl 4-nitroanilide
0.5
N-acetyl-L-leucyl 4-nitroanilide
Thermococcus onnurineus
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pH 7.5, 80°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
41 - 47
L-leucyl 4-nitroanilide
12218
55 - 220
L-methionyl 4-nitroanilide
15290
0.45
N-acetyl-L-leucyl 4-nitroanilide
Thermococcus onnurineus
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pH 7.5, 80°C
10402
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
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pH 6.5: about 50% of maximal activity, pH 8.5: about 70% of maximal activity (HEPES buffer), about 40% of maximal activity (Tris-HCl buffer)
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
assay at; assay at; assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60 - 100
60°C: about 60% of maximal activity, 80°C: about 90% of maximal activity, 100°C: about 95% of maximal activity
70 - 100
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70°C: about 35% of maximal activity, 90-100°C: optimum
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36470
4 * 36470, calculated from sequence
37360
2 * 37360, calculated from sequence
37900
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x * 37900, His-tagged enzyme, SDS-PAGE
37950
14 * 37950, calculated from sequence
71000
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dimeric form prevalent in presence of 0.1 mM Co2+, gel filtration
76100
MALDI-TOF mas spectrometry
148000
isoenzyme TNA1-DAP3, MALDI-TOF mass spectrometry
539600
MALDI-TOF mass spectrometry
600000
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oligomeric form prevalent in absence of Co2+, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 37900, His-tagged enzyme, SDS-PAGE
dimer
2 * 37360, calculated from sequence
tetradecamer
14 * 37950, calculated from sequence
tetramer
4 * 36470, calculated from sequence
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli