Information on EC 3.4.22.7 - asclepain

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The expected taxonomic range for this enzyme is: Asclepiadeae

EC NUMBER
COMMENTARY
3.4.22.7
-
RECOMMENDED NAME
GeneOntology No.
asclepain
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Similar to that of papain
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
endopeptidase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
asclepain c
-
-
asclepain cI
-
-
asclepain cI
B7VF64
-
asclepain cII
-
-
asclepain cII
B7VF65
-
asclepain f
B5BLP0
synthesized as preproenzyme
papain-like endopeptidase
-
-
papain-like protease
B7VF64, B7VF65
-
CAS REGISTRY NUMBER
COMMENTARY
37288-80-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
fragment; scarlet milkweed
B7VF64, B7VF65
UniProt
Manually annotated by BRENDA team
strain H.B.K., ten asclepain forms
-
-
Manually annotated by BRENDA team
Asclepias glaucescens H.B.K.
strain H.B.K., ten asclepain forms
-
-
Manually annotated by BRENDA team
Asclepias sp.
-
-
-
Manually annotated by BRENDA team
Asclepias sp.
asclepain A4 and B1
-
-
Manually annotated by BRENDA team
asclepain A1, A2, A3, A4, A5, and asclepain B1, B2, B3, B4 and B5
-
-
Manually annotated by BRENDA team
asclepain A3 and B5
-
-
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
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
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
casein + H2O
?
show the reaction diagram
-
about 50% degradation after 6 h
-
-
?
casein + H2O
?
show the reaction diagram
Asclepias glaucescens H.B.K.
-
-
-
-
?
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
-
-
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
B7VF64, B7VF65
very low activity
-
-
?
L-pyroglutamyl-L-phenylalanyl-L-leucine-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
N-acetyl-L-tryptophanamide + H2O
N-acetyl-L-Trp + NH3
show the reaction diagram
Asclepias glaucescens, Asclepias glaucescens H.B.K.
-
-
-
-
?
N-alpha-benzyloxycarbonyl-Gly-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Ala-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Ala + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Asn-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Asn + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Asp-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Asp + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
asclepain cII has highest endoesterolytic activity using N-alpha-benzyloxycarbonyl-amino acid p-nitrophenyl esters with L-Asp as amino acid
-
-
?
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Gln + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
N-alpha-benzyloxycarbonyl-L-Tyr-4-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-L-Tyr + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
-
-
-
?
N-Benzoyl-L-Arg ethyl ester + H2O
N-Benzoyl-L-Arg + ethanol
show the reaction diagram
Asclepias glaucescens, Asclepias glaucescens H.B.K.
-
-
-
-
?
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
-
-
-
N-carbobenzoxy-L-Gln 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Gln + 4-nitrophenol
show the reaction diagram
-
-
-
-
-
N-carbobenzoxy-L-Ile 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Ile + 4-nitrophenol
show the reaction diagram
-
59% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
-
-
-
N-Carbobenzoxy-L-Lys 4-nitrophenyl ester + H2O
N-Carbobenzoxy-L-Lys + 4-nitrophenol
show the reaction diagram
-
78% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
-
-
-
N-carbobenzoxy-L-Tyr 4-nitrophenyl ester + H2O
N-carbobenzoxy-L-Tyr + 4-nitrophenol
show the reaction diagram
-
51% of the activity with N-carbobenzoxy-L-Gln 4-nitrophenyl ester
-
-
-
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
-
-
-
-
?
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.
-
-
-
-
?
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
-
about 25% activity compared to Nalpha-carbobenzoxy-L-Ala-4-nitrophenyl ester
-
-
?
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
-
-
?
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
-
-
?
Nalpha-CBZ-L-Gln-4-nitrophenyl ester + H2O
Nalpha-CBZ-L-Gln + 4-nitrophenol
show the reaction diagram
B7VF64, B7VF65
when assayed on N-alpha-CBZ-amino acid-4-nitrophenyl esters, the enzyme exhibits higher preference for the glutamine derivative
-
-
?
oxidized insulin B chain + H2O
?
show the reaction diagram
Asclepias sp.
-
detection of cleavage points
-
-
?
pGlu-Phe-Leu-p-nitroanilide + H2O
pGlu-Phe-Leu + p-nitroaniline
show the reaction diagram
B5BLP0
-
-
-
?
Z-Arg-p-nitroanilide + H2O
Z-Arg + p-nitroaniline
show the reaction diagram
-
-
-
-
?
L-pyroglutamyl-Phe-Leu-p-nitroanilide + H2O
L-pyroglutamyl-Phe-Leu + p-nitroaniline
show the reaction diagram
-
-
-
-
?
additional information
?
-
B7VF64, B7VF65
asclepain cI is the main active enzyme, but shows lower specific activity than asclepain cII, which is the minor proteolytic component in the latex, overview
-
-
-
additional information
?
-
B7VF64, B7VF65
asclepain cII is the minor proteolytic component in the latex, but shows higher specific activity than asclepain cI, the main active enzyme, overview
-
-
-
additional information
?
-
B7VF64, B7VF65
the peptide PFLNA is a poor substrate for asclepain cII
-
-
-
additional information
?
-
B7VF64, B7VF65
no activity towards N-alpha-benzyloxycarbonyl-L-Val-4-nitrophenyl ester, N-alpha-benzyloxycarbonyl-L-Leu-4-nitrophenyl ester, N-alpha-benzyloxycarbonyl-L-Ile-4-nitrophenyl ester, and N-alpha-benzyloxycarbonyl-L-Lys-4-nitrophenyl ester
-
-
-
additional information
?
-
-
ascpelain f shows low milk clotting activity, no activity towards Nalpha-carbobenzoxy-L-Leu-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Asn-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Trp-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Val-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Asp-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-lys-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Phe-4-nitrophenyl ester, Nalpha-carbobenzoxy-L-Pro-4-nitrophenyl ester, and Nalpha-carbobenzoxy-L-Ile-4-nitrophenyl ester
-
-
-
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
casein + H2O
?
show the reaction diagram
-
about 50% degradation after 6 h
-
-
?
additional information
?
-
B7VF64, B7VF65
asclepain cI is the main active enzyme, but shows lower specific activity than asclepain cII, which is the minor proteolytic component in the latex, overview
-
-
-
additional information
?
-
B7VF64, B7VF65
asclepain cII is the minor proteolytic component in the latex, but shows higher specific activity than asclepain cI, the main active enzyme, overview
-
-
-
additional information
?
-
-
ascpelain f shows low milk clotting activity
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(2S,3S)-3-(N-[(S)-1-[N-(4-guanidinobutyl)carbamoyl]3-methylbutyl]carbamoyl)oxirane-2-carboxylic acid
B7VF64, B7VF65
i.e. E-64, complete immediate and irreversible inhibition at 10 mM after 30 min incubation
E-64
B7VF64, B7VF65
;
NaCl
-
1 M, 70% loss of activity
phenylmethylsulfonyl fluoride
B7VF64, B7VF65
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
-
-
iodoacetic acid
-
-
additional information
B7VF64, B7VF65
1,10-phenantroline and pepstatine do not affect the activity of asclepain cII; activity of asclepain cII is inhibited by cysteine proteases inhibitors like E-64, but not by any other protease inhibitors such as 1,10-phenantroline, phenylmethanesulfonyl fluoride, and pepstatine
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.73
-
L-pyroglutamyl-L-phenylalanyl-L-leucine-4-nitroanilide
-
pH 8.5, 42C, Lineweaver-Burk equation
0.1634
-
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
B7VF64, B7VF65
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
0.0097
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A5
0.0105
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A2
0.0114
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A1
0.0122
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A4
0.0126
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B3
0.0142
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A3
0.0248
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B1
0.027
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B4
0.0272
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B5
0.0389
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B2
0.163
-
Nalpha-CBZ-L-Gln-4-nitrophenyl ester
B7VF64, B7VF65
pH 8.5, 42C
0.82
-
L-pyroglutamyl-L-phenylalanyl-L-leucine-4-nitroanilide
-
pH 8.5, 42C, Eddie-Hofstee equation
additional information
-
additional information
Asclepias sp.
-
Km-values for hydrolysis of aryl mesyl glycinates
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
121.5
-
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
B7VF64, B7VF65
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
22.3
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A5
25.8
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B3
33.8
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B4
34.6
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B1
42.9
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A3
46.2
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A2
47.6
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B5
49.2
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain B2
54.7
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A1
59.6
-
N-carbobenzoxy-Lys p-nitrophenyl ester
-
asclepain A4
121.5
-
Nalpha-CBZ-L-Gln-4-nitrophenyl ester
B7VF64, B7VF65
pH 8.5, 42C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
740000
-
N-alpha-benzyloxycarbonyl-L-Gln-4-nitrophenyl ester
B7VF64, B7VF65
at 37C, in 0.1 M Tris-HCl buffer (pH 8.0) containing 2 mM EDTA and 25 mM cysteine
302627
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12.8
-
B7VF64, B7VF65
purified asclepain cII
480
-
-
substrate Z-Arg-p-nitroanilide, immobilized enzyme
4100
-
-
asclepain A3
6300
-
-
ascepain B5
10880
-
-
pH 8.5, 42C
additional information
-
B7VF64, B7VF65
the enzyme has a specific activity of 1.0586 units/mg in crude extract and of 12.836 units/mg after 12.1 fold purification, using casein as substrate, one arbitrary enzyme unit is defined as the amount of enzyme that produces an increase of one absorbance unit per minute
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
8
-
hydrolysis of N-benzoyl-L-Arg ethyl ester, asclepain Ag7
7
7.5
-
hydrolysis of casein, asclepain B5
7.5
8.5
-
hydrolysis of casein, asclepain A3
8.5
-
B7VF64, B7VF65
-
9.4
10.2
B7VF64, B7VF65
asclepain cII
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
9
-
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
B7VF64, B7VF65
asclepain cII shows maximum activity within the range of pH 9.4 and 10.2 with casein as substrate
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
42
-
B7VF64, B7VF65
assay at
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
9.3
-
-
higher than, isoelectric focusing
9.3
-
B7VF64, B7VF65
above, asclepain cII, isoelectric focusing; isoelectric focusing
9.78
-
-, B7VF64, B7VF65
calculated
9.82
-
-, B7VF64, B7VF65
calculated
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
B7VF64, B7VF65
extracted from petioles; extracted from petioles; latex extracted from petioles
Manually annotated by BRENDA team
-, B7VF64, B7VF65
;
Manually annotated by BRENDA team
Asclepias glaucescens H.B.K.
-
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
B7VF64, B7VF65
asclepain cI; asclepain cII
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
19000
-
-
asclepain B5, gel filtration
21800
-
-
asclepain Ag3, gel filtration
22500
-
-
asclepain Ag6, gel filtration
22600
-
-
asclepain Ag7, gel filtration
22700
-
-
asclepain Ag8, gel filtration
23500
-
B7VF64, B7VF65
SDS-PAGE
23590
-
B7VF64, B7VF65
MALDI-TOF mass spectrometry
24000
-
-
asclepain A3, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 23200, mass spectrometry
?
B7VF64, B7VF65
x * 23500, asclepain cII, SDS-PAGE, x * 23590, asclepain cII, mass spectrometry
?
-, B7VF64, B7VF65
x * 23057, calculated; x * 23513, calculated
?
B5BLP0
x * 23406, calculated
?
-
x * 21000, asclepain A, SDS-PAGE; x * 23000, asclepain B, SDS-PAGE
monomer
-
1 * 23000, carboxymethylated asclepain A3 and carboxymethylated asclepain B5, SDS-PAGE
monomer
-
1 * 23300, asclepain Ag7, SDS-PAGE; 1 * 23400, asclepain Ag3, SDS-PAGE; 1 * 24200, asclepain Ag8, SDS-PAGE; 1 * 24400, asclepain Ag6, SDS-PAGE
monomer
Asclepias glaucescens H.B.K.
-
1 * 23300, asclepain Ag7, SDS-PAGE; 1 * 23400, asclepain Ag3, SDS-PAGE; 1 * 24200, asclepain Ag8, SDS-PAGE; 1 * 24400, asclepain Ag6, SDS-PAGE
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no modification
-
no bound carbohydrate
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8.5
-
-
2 h at 37C, stable, 4 h, 30% loss of activity
10
-
B7VF64, B7VF65
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
additional information
-
-
unstable in acid or alkali
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
2 h at pH 8.5, stable, 4 h, 30% loss of activity
40
-
-
2 h, stable
50
70
B7VF64, B7VF65
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
50
-
-
2 h, stable
50
-
B7VF64, B7VF65
2 h, asclepain cII, 30% remaining activity
60
-
-
2 h, stable
70
-
-
40 min, complete inactivation
additional information
-
B7VF64, B7VF65
purified asclepain cII shows poor thermostability, profile, overview
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cation exchange chromatography; cation exchange chromatography
-, B7VF64, B7VF65
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
B7VF64, B7VF65
-
Asclepias sp.
-
partial purification
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Pichia pastoris
B5BLP0
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
food industry
-
asclepain f is less adequate as coagulant in cheesemaking