Information on EC 3.4.17.B4 - Thermococcus onnurineus carboxypeptidase

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The expected taxonomic range for this enzyme is: Thermococcus onnurineus

EC NUMBER
COMMENTARY hide
3.4.17.B4
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
Thermococcus onnurineus carboxypeptidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Broad substrate specificity with a preference for basic, aliphatic, and aromatic C-terminal amino acids. No activity with N-benzyloxycarbonyl-Ala-Asp, N-benzyloxycarbonyl-Ala-Asn, N-benzyloxycarbonyl-Ala-Pro, N-benzyloxycarbonyl-Ala-His.
show the reaction diagram
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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
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro + L-phenylalanine
show the reaction diagram
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-
?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe + L-histidine
show the reaction diagram
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?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His + L-leucine
show the reaction diagram
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-
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?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu + L-leucine
show the reaction diagram
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?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu + L-valine
show the reaction diagram
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-
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?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val + L-tyrosine
show the reaction diagram
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-
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?
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr-Ser + H2O
Ac-Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr + L-serine
show the reaction diagram
release of up to seven residues from the C-terminus
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?
N-benzyloxycarbonyl-Ala-Ala + H2O
N-benzyloxycarbonyl-Ala + L-alanine
show the reaction diagram
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-
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?
N-benzyloxycarbonyl-Ala-Arg + H2O
N-benzyloxycarbonyl-Ala + L-arginine
show the reaction diagram
compared to the activity of Pyrococcus furiosus carboxypeptidase the catalytic efficiency is of Thermococcus onnurineus carboxypeptidase is 80-fold lower
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?
N-benzyloxycarbonyl-Ala-Leu + H2O
N-benzyloxycarbonyl-Ala + L-leucine
show the reaction diagram
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?
N-benzyloxycarbonyl-Ala-Met + H2O
N-benzyloxycarbonyl-Ala + L-methionine
show the reaction diagram
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?
N-benzyloxycarbonyl-Ala-Tyr + H2O
N-benzyloxycarbonyl-Ala + L-tyrosine
show the reaction diagram
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?
N-benzyloxycarbonyl-Ala-Val + H2O
N-benzyloxycarbonyl-Ala + L-valine
show the reaction diagram
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?
additional information
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no activity with N-benzyloxycarbonyl-Ala-Asp, N-benzyloxycarbonyl-Ala-Asn, N-benzyloxycarbonyl-Ala-Pro, N-benzyloxycarbonyl-Ala-His
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
0.2 mM, stimulates activity, can not be replaced by other divalent metal ions, no activation by Zn2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
0.2 mM, about 90% loss of activity, the inhibitory effect is not overcome by the presence of Co2+
EDTA
1 mM, complete inactivation
Fe2+
0.2 mM, about 70% loss of activity, the inhibitory effect is not overcome by the presence of Co2+
Ni2+
0.2 mM, about 75% loss of activity, the inhibitory effect is not overcome by the presence of Co2+
Zn2+
0.2 mM, about 90% loss of activity, the inhibitory effect is not overcome by the presence of Co2+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.4
N-benzyloxycarbonyl-Ala-Arg
pH 6.5, 70-80C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11.6
N-benzyloxycarbonyl-Ala-Arg
Thermococcus onnurineus
Q2QC89
pH 6.5, 70-80C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.3
N-benzyloxycarbonyl-Ala-Arg
Thermococcus onnurineus
Q2QC89
pH 6.5, 70-80C
42211
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
substrate: N-benzyloxycarbonyl-Ala-Arg
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3 - 7.5
pH 5.3: about 80% of maximal activity, pH 7.5: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
70 - 80
substrate: N-benzyloxycarbonyl-Ala-Arg
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50 - 90
50C: about 50% of maximal activity, 90C: about 60% of maximal activity
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
61000
2 * 61000, SDS-PAGE
135000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
2 * 61000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
pH 6.5, 0.2 mM Co2+, t1/2: 84 min, t1/2 in absence of Co2+ is 7.4 min
90
pH 6.5, 0.2 mM Co2+, t1/2: 8 min, t1/2 in absence of Co2+ is 5 min
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glycerol
12.5%, does not protect from inactivation at either temperature
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overexpression in Escherichia coli