Information on EC 3.4.24.25 - vibriolysin

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.24.25
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RECOMMENDED NAME
GeneOntology No.
vibriolysin
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Preferential cleavage of bonds with bulky hydrophobic groups in P2 and P1'. Phe at P1' is the most favoured residue, which distinguished this enzyme from thermolysin
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY hide
58500-42-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
precursor; strain 643
SwissProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-Furanacryloyl-Gly-(2-hydroxyisohexanoic-acid)-NH2 + H2O
?
show the reaction diagram
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-
-
-
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3-(2-furylacryloyl)glycyl-L-leucine + H2O
?
show the reaction diagram
aminopeptidase + H2O
?
show the reaction diagram
-
vibriolysin activates aminopeptidase
-
-
?
azocasein + H2O
?
show the reaction diagram
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-
-
?
Benzoyl-Gly-(2-hydroxyisohexanoic acid)-Ala + H2O
?
show the reaction diagram
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-
-
-
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Benzoyl-Gly-(2-phenyllactic acid)-Ala + H2O
?
show the reaction diagram
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-
-
-
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Benzoyl-Gly-(2-phenyllactic acid)-Phe + H2O
?
show the reaction diagram
-
-
-
-
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Benzoyl-Gly-Leu-Ala + H2O
?
show the reaction diagram
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-
-
-
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Benzoyl-Gly-Phe-Gly + H2O
?
show the reaction diagram
-
-
-
-
-
Benzoyl-Gly-Phe-Phe + H2O
?
show the reaction diagram
-
-
-
-
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Benzoyl-Phe-(2-phenyllactic acid)-Ala + H2O
?
show the reaction diagram
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-
-
-
-
Benzoyl-Phe-Phe-Ala + H2O
?
show the reaction diagram
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-
-
-
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Benzoyl-Phe-Phe-Gly + H2O
?
show the reaction diagram
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-
-
-
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Benzyloxycarbonyl-Ala-Phe-NH2 + H2O
?
show the reaction diagram
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-
-
-
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Benzyloxycarbonyl-Gly-Ala-NH2 + H2O
?
show the reaction diagram
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-
-
-
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Benzyloxycarbonyl-Gly-Ile-NH2 + H2O
?
show the reaction diagram
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-
-
-
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Benzyloxycarbonyl-Gly-Leu-NH2 + H2O
?
show the reaction diagram
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-
-
-
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Benzyloxycarbonyl-Gly-norvaline-NH2 + H2O
?
show the reaction diagram
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-
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Benzyloxycarbonyl-Gly-Phe-NH2 + H2O
?
show the reaction diagram
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-
-
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Benzyloxycarbonyl-Gly-Tyr-NH2 + H2O
?
show the reaction diagram
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-
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Benzyloxycarbonyl-Gly-Val-NH2 + H2O
?
show the reaction diagram
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-
-
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Benzyloxycarbonyl-Leu-Phe-NH2 + H2O
?
show the reaction diagram
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-
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Benzyloxycarbonyl-Ser-Phe-NH2 + H2O
?
show the reaction diagram
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casein + H2O
?
show the reaction diagram
Collagen + H2O
?
show the reaction diagram
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from lumpfish skin
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-
?
furylacryloyl-Gly-Leu-NH2 + H2O
furylacryloyl-Gly + Leu-NH2
show the reaction diagram
Furylacryloyl-Gly-Phe-Gly-NH2 + H2O
?
show the reaction diagram
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Gelatin + H2O
?
show the reaction diagram
Urea-denatured hemoglobin + H2O
?
show the reaction diagram
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additional information
?
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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
aminopeptidase + H2O
?
show the reaction diagram
-
vibriolysin activates aminopeptidase
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-
?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Calcium
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contains 1.7 mol of calcium per mol of enzyme, after 48 h of dialysis, not essential for activity, Ca2+ likely plays a structural role
Zn2+
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a zinc metalloprotease
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
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1 mM, 73% loss of activity
3-Phenyl-1-propanol
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3-Phenylpropionyl-L-Phe
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pH-dependence of inhibition
3-Phenylpropionyl-Phe-NH2
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Acetohydroxamate
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Aminoacyl hydroxamates
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Chloroacetyl-N-hydroxy-L-Phe-L-Ala-L-Ala
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EDTA
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1 mM, 16% loss of activity
Glycine hydroxamate
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L-Leucine hydroxamate
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L-phenylalanine
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L-phenylalanine hydroxamate
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N-tert-Butoxycarbonyl-L-Phe hydroxamate
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N-[(1-carboxy-3-phenylpropyl)-L-phenylalanyl-alpha-L-asparagine]
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Peptides containing a phenylalanyl residue
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Phe-Ala-Gly
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Phe-Ala-Gly-Gly
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Phosphoryl-L-Phe-Gly-Gly
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Phosphoryl-Phe-Gly-Gly
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.8
Benzyloxycarbonyl-Gly-Ala-NH2
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2.1
Benzyloxycarbonyl-Gly-Ile-NH2
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2.2
Benzyloxycarbonyl-Gly-Leu-NH2
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0.2
benzyloxycarbonyl-Gly-Phe-Gly-NH2
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0.34
Benzyloxycarbonyl-Gly-Phe-NH2
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1.2
Benzyloxycarbonyl-Gly-Tyr-NH2
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1.9
Benzyloxycarbonyl-Gly-Val-NH2
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2.6
benzyloxycarbonyl-norvaline-NH2
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3.5
Furylacryloyl-Gly-Leu-NH2
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0.32
furylacryloyl-Gly-Phe-NH2
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additional information
additional information
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overview: effect of residues at P1', P1'' or P2' on Km, influence of the identity of residues remote from scissile bond
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
Benzyloxycarbonyl-Gly-Ile-NH2
Vibrio proteolyticus
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25
Benzyloxycarbonyl-Gly-Leu-NH2
Vibrio proteolyticus
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4
Benzyloxycarbonyl-Gly-norvaline-NH2
Vibrio proteolyticus
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1150
benzyloxycarbonyl-Gly-Phe-Gly-NH2
Vibrio proteolyticus
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0.4 - 584
Benzyloxycarbonyl-Gly-Phe-NH2
38
Benzyloxycarbonyl-Gly-Tyr
Vibrio proteolyticus
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0.5
Benzyloxycarbonyl-Gly-Tyr-NH2
Vibrio proteolyticus
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additional information
additional information
Vibrio proteolyticus
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overview: effect of residues at P1', P1'' or P2' on turnover number, influence of the identity of residues remote from scissile bond
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.32 - 2.55
3-(2-furylacryloyl)glycyl-L-leucine
163627
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1736
recombinant enzyme, 37C, pH 7.4
2020
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substrate casein, 30C
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
using casein as substrate, at 60C
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
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or 37C, assay at
37
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or 25C, assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
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35% residual activity
40 - 70
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudoalteromonas sp. (strain SM9913)
Pseudoalteromonas sp. (strain SM9913)
Pseudoalteromonas sp. (strain SM9913)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34800
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Aeromonas proteolytica, sedimentation equilibrium, sedimentation velocity-diffusion experiments
45000
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x * 45000, pro-vibriolysin, SDS-PAGE, x * 35000, vibriolysin, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 45000, pro-vibriolysin, SDS-PAGE, x * 35000, vibriolysin, SDS-PAGE
monomer
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1 * 35000 (about), Aeromonas proteolytica, sedimentation equilibrium in presence of guanidine-HCl
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 11
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10C, stable for more than 1 week
31109
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
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30 min, 60% residual activity
55 - 64
70
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heat treatment (70C) effectively inactivates vibriolysin
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Lyophilization and reconstitution results in approximately 20% loss of activity
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, Tricine or Tris buffer, stable for several years
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation and DEAE-Sepharose column chromatography
recombinant processed 35 kDa vibriolysin from Escherichia coli by anion exchange chromatography and gel filtration to homogeneity
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recombinant protein
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3) cells. Vibriolysin is not expressed as an active form in Escherichia coli because of the strong inhibitory effect of its N-terminal propeptide. The vibriolysin NprV-R is expressed extracellularly as an active form in Escherichia coli
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expression in Escherichia coli
expression of pro-vibriolysin in Escherichia coli and cleavage of the N-terminal propeptide
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
isolation of a vibriolysin mutant by in vivo random mutagenensis. contrary to wild-type, the mutant is expressed as active mature vibriolysin in Escherichia coli. The N-terminal propeptide of the engineered enzyme is processed and degraded, confirming that the propeptide inhibits the mature enzyme. Two mutations result in the substitution of stop codon for Trp at position 11 in the signal peptide and of Val for Ala at position 183 in the N-terminal propeptide