Information on EC 3.4.22.7 - asclepain

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The expected taxonomic range for this enzyme is: Asclepiadeae

EC NUMBER
COMMENTARY hide
3.4.22.7
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RECOMMENDED NAME
GeneOntology No.
asclepain
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Similar to that of papain
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
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endopeptidase
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CAS REGISTRY NUMBER
COMMENTARY hide
37288-80-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain H.B.K., ten asclepain forms
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Manually annotated by BRENDA team
strain H.B.K., ten asclepain forms
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
Asclepias sp.
-
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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asclepain cII present in the latex of Asclepias curassavica is found to accelerate the blood coagulation process. Results reveal that asclepain cII present in the latex is responsible for its thrombin like activity
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
benzoyl-L-Arg-p-nitroanilide + H2O
benzoyl-L-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
insulin B-chain + H2O
Phe-Val-Asn + Ser-His-Leu-Val-Glu-Ala-Leu + Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-CysA-Gly-Glu + Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala + Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala + Gly-Phe-Phe-Tyr-Thr
show the reaction diagram
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-
14% Phe-Val-Asn + 22% Ser-His-Leu-Val-Glu-Ala-Leu + 15% Leu-Val-Glu-Ala-Leu-Tyr-Leu-Val-CysA-Gly-Glu + 10% Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala + 10% Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala + 13% Gly-Phe-Phe-Tyr-Thr
?
L-pyroglutamyl-L-phenylalanyl-L-leucine 4-nitroanilide + H2O
L-pyroglutamyl-L-phenylalanyl-L-leucine + 4-nitroaniline
show the reaction diagram
very low activity
-
-
?
L-pyroglutamyl-L-phenylalanyl-L-leucine-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
L-pyroglutamyl-Phe-Leu-p-nitroanilide + H2O
L-pyroglutamyl-Phe-Leu + p-nitroaniline
show the reaction diagram
-
-
-
-
?
N-acetyl-L-tryptophanamide + H2O
N-acetyl-L-Trp + NH3
show the reaction diagram
N-alpha-benzyloxycarbonyl-Gly-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Ala-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Ala + 4-nitrophenol
show the reaction diagram
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Asn-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Asn + 4-nitrophenol
show the reaction diagram
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Asp-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Asp + 4-nitrophenol
show the reaction diagram
asclepain cII has highest endoesterolytic activity using N-alpha-benzyloxycarbonyl-amino acid p-nitrophenyl esters with L-Asp as amino acid
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-
?
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Gln + 4-nitrophenol
show the reaction diagram
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Tyr-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Tyr + 4-nitrophenol
show the reaction diagram
-
-
-
?
N-Benzoyl-L-Arg ethyl ester + H2O
N-Benzoyl-L-Arg + ethanol
show the reaction diagram
N-carbobenzoxy-Gly p-nitrophenyl ester + H2O
N-carbobenzoxy-Gly + p-nitrophenol
show the reaction diagram
-
-
-
-
?
N-carbobenzoxy-L-Ala 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Ala + 4-nitrophenol
show the reaction diagram
-
47% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
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-
N-carbobenzoxy-L-Gln 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Gln + 4-nitrophenol
show the reaction diagram
-
-
-
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N-carbobenzoxy-L-Ile 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Ile + 4-nitrophenol
show the reaction diagram
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59% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
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N-Carbobenzoxy-L-Lys 4-nitrophenyl ester + H2O
N-Carbobenzoxy-L-Lys + 4-nitrophenol
show the reaction diagram
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78% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
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N-carbobenzoxy-L-Tyr 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Tyr + 4-nitrophenol
show the reaction diagram
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51% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
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N-carbobenzoxy-Leu p-nitrophenyl ester + H2O
N-carbobenzoxy-Leu + p-nitrophenol
show the reaction diagram
-
-
-
-
?
N-carbobenzoxy-Lys p-nitrophenyl ester + H2O
N-carbobenzoxy-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
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?
N-carbobenzoxy-Tyr p-nitrophenyl ester + H2O
N-carbobenzoxy-Tyr + p-nitrophenol
show the reaction diagram
-
-
-
-
?
N-methanesulfonylglycine 3-bromophenyl ester + H2O
N-methanesulfonylglycine + 3-bromophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 3-chlorophenyl ester + H2O
N-methanesulfonylglycine + 3-chlorophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 3-fluorophenyl ester + H2O
N-methanesulfonylglycine + 3-fluorophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 3-methylphenyl ester + H2O
N-methanesulfonylglycine + 3-methylphenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 3-nitrophenyl ester + H2O
N-methanesulfonylglycine 3-nitrophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 3-tert-butylphenyl ester + H2O
N-methanesulfonylglycine + 3-tert-butylphenol
show the reaction diagram
Asclepias sp.
-
-
-
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?
N-methanesulfonylglycine 4-bromophenyl ester + H2O
N-methanesulfonylglycine + 4-bromophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 4-chlorophenyl ester + H2O
N-methanesulfonylglycine + 4-chlorophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 4-fluorophenyl ester + H2O
N-methanesulfonylglycine + 4-fluorophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 4-methylphenyl ester + H2O
N-methanesulfonylglycine + 4-methylphenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
N-methanesulfonylglycine 4-nitrophenyl ester + H2O
N-methanesulfonylglycine + 4-nitrophenol
show the reaction diagram
Asclepias sp.
-
-
-
-
?
Nalpha-carbobenzoxy-Gly-4-nitrophenyl ester + H2O
Nalpha-carbobenzoxy-Gly + 4-nitrophenol
show the reaction diagram
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about 25% activity compared to Nalpha-carbobenzoxy-L-Ala-4-nitrophenyl ester
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?
Nalpha-carbobenzoxy-L-Ala-4-nitrophenyl ester + H2O
Nalpha-carbobenzoxy-L-Ala + 4-nitrophenol
show the reaction diagram
-
100% activity
-
-
?
Nalpha-carbobenzoxy-L-Gln-4-nitrophenyl ester + H2O
Nalpha-carbobenzoxy-L-Gln + 4-nitrophenol
show the reaction diagram
-
about 85% activity compared to Nalpha-carbobenzoxy-L-Ala-4-nitrophenyl ester
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-
?
Nalpha-carbobenzoxy-L-Tyr-4-nitrophenyl ester + H2O
Nalpha-carbobenzoxy-L-Tyr + 4-nitrophenol
show the reaction diagram
-
about 20% activity compared to Nalpha-carbobenzoxy-L-Ala-4-nitrophenyl ester
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?
Nalpha-CBZ-L-Gln-4-nitrophenyl ester + H2O
Nalpha-CBZ-L-Gln + 4-nitrophenol
show the reaction diagram
when assayed on N-alpha-CBZ-amino acid-4-nitrophenyl esters, the enzyme exhibits higher preference for the glutamine derivative
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-
?
oxidized insulin B chain + H2O
?
show the reaction diagram
Asclepias sp.
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detection of cleavage points
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-
?
pGlu-Phe-Leu-p-nitroanilide + H2O
pGlu-Phe-Leu + p-nitroaniline
show the reaction diagram
-
-
-
?
Z-Arg-p-nitroanilide + H2O
Z-Arg + p-nitroaniline
show the reaction diagram
-
-
-
-
?
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
casein + H2O
?
show the reaction diagram
-
about 50% degradation after 6 h
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-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3S)-3-(N-[(S)-1-[N-(4-guanidinobutyl)carbamoyl]3-methylbutyl]carbamoyl)oxirane-2-carboxylic acid
i.e. E-64, complete immediate and irreversible inhibition at 10 mM after 30 min incubation
iodoacetic acid
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-
NaCl
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1 M, 70% loss of activity
phenylmethylsulfonyl fluoride
incubation with 100 mM phenylmethylsulfonyl fluoride for 30 min decreases activity to 63% of the initial value, but inhibition is partially reverted when 12 mM cysteine is added to the reaction mixture
Sodium tetrathionate
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-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.73 - 0.82
L-pyroglutamyl-L-phenylalanyl-L-leucine-4-nitroanilide
0.1634
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
0.0097 - 0.0389
N-carbobenzoxy-Lys p-nitrophenyl ester
0.163
Nalpha-CBZ-L-Gln-4-nitrophenyl ester
pH 8.5, 42C
additional information
additional information
Asclepias sp.
-
Km-values for hydrolysis of aryl mesyl glycinates
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
121.5
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
Asclepias curassavica
B7VF64, B7VF65
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
22.3 - 59.6
N-carbobenzoxy-Lys p-nitrophenyl ester
121.5
Nalpha-CBZ-L-Gln-4-nitrophenyl ester
Asclepias curassavica
B7VF64, B7VF65
pH 8.5, 42C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
740000
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
Asclepias curassavica
B7VF64, B7VF65
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
40629
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12.8
purified asclepain cII
480
-
substrate Z-Arg-p-nitroanilide, immobilized enzyme
4100
-
asclepain A3
6300
-
ascepain B5
10880
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pH 8.5, 42C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
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hydrolysis of N-benzoyl-L-Arg ethyl ester, asclepain Ag7
7 - 7.5
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hydrolysis of casein, asclepain B5
7.5 - 8.5
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hydrolysis of casein, asclepain A3
9.4 - 10.2
asclepain cII
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
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pH 5.0: about 70% of maximal activity, pH 9.0: about 30% of maximal activity, hydrolysis of N-benzoyl-L-Arg ethyl ester, asclepain Ag7
9.4 - 10.2
asclepain cII shows maximum activity within the range of pH 9.4 and 10.2 with casein as substrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
asclepain cI; asclepain cII
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
19000
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asclepain B5, gel filtration
21800
-
asclepain Ag3, gel filtration
22500
-
asclepain Ag6, gel filtration
22600
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asclepain Ag7, gel filtration
22700
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asclepain Ag8, gel filtration
23590
MALDI-TOF mass spectrometry
24000
-
asclepain A3, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no modification
-
no bound carbohydrate
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
-
2 h at 37C, stable, 4 h, 30% loss of activity
670802
10
1 h, asclepain cII, about 95% of the residual activity; asclepain cII shows a high stability at pH 10.0 retaining about 95% of the residual activity after 1 h
695296
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
2 h at pH 8.5, stable, 4 h, 30% loss of activity
40
-
2 h, stable
50 - 70
asclepain cII exhibits poor thermostability, as only 30% of its activity is retained after 2 h of incubation at 50C, at 60-70C the loss of activity is rapid
60
-
2 h, stable
70
-
40 min, complete inactivation
additional information
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cation exchange chromatography; cation exchange chromatography
native asclepain cII 12.1fold to homogeneity by cation exchange chromatography, separation from asclepain cI; separation of asclepain cI from asclepain cII; SP-Sepharose column chromatography, gel filtration
partial purification
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
food industry
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asclepain f is less adequate as coagulant in cheesemaking