BRENDA - Enzyme Database

Characterization of a thermostable lysine-specific metalloendopeptidase from the fruiting bodies of a basidiomycete, Grifola frondosa

Nonaka, T.; Ishikawa, H.; Tsumuraya, Y.; Hashimoto, Y.; Dohmae, N.; Takio, K.; J. Biochem. 118, 1014-1020 (1995)

Data extracted from this reference:

Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.4.24.20
1,10-phenanthroline
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
2,2'-bipyridine
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
2-mercaptoethanol
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
6-aminohexanoic acid
caseinolytic activity
Grifola frondosa
3.4.24.20
agmatine
caseinolytic activity
Grifola frondosa
3.4.24.20
antipain
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Ba2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Ca2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Co2+
0.1 mM, caseinolytic activity
Grifola frondosa
3.4.24.20
Cu2+
1 mM, caseinolytic activity
Grifola frondosa
3.4.24.20
dithiothreitol
caseinolytic activity; strong
Grifola frondosa
3.4.24.20
DL-homoarginine
caseinolytic activity
Grifola frondosa
3.4.24.20
EDTA
fully restorable by Ca2+; fully restorable by Zn2+, Mn2+, Co2+; strong, caseinolytic activity
Grifola frondosa
3.4.24.20
Hg2+
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
L-aminobutyric acid
caseinolytic activity
Grifola frondosa
3.4.24.20
L-arginine
caseinolytic activity
Grifola frondosa
3.4.24.20
L-lysine
caseinolytic activity
Grifola frondosa
3.4.24.20
L-Lysine ethylester
caseinolytic activity
Grifola frondosa
3.4.24.20
L-ornithine
caseinolytic activity
Grifola frondosa
3.4.24.20
Mg2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
additional information
no inhibition (of caseinolytic activity) by chymostatin, iodoacetate, leupeptin; PMSF; zinc acetate, elastatinal, Lys-Lys, 4 M urea
Grifola frondosa
3.4.24.20
p-chloromercuribenzoate
not
Grifola frondosa
3.4.24.20
phosphoramidon
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
tosyl-Lys chloromethyl ketone
not
Grifola frondosa
3.4.24.20
tosyl-Phe chloromethyl ketone
weak, caseinolytic activity
Grifola frondosa
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.4.24.20
cytosol
-
Grifola frondosa
5829
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.4.24.20
Ca2+
activation, zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Co2+
activation, zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Mn2+
activation, 0.1 mM, native enzyme and zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Zinc
atomic absorption spectroscopy; requirement, 1 atom per molecule of native enzyme; Zn2+ activates zinc-depleted apoenzyme
Grifola frondosa
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.4.24.20
20000
-
x * 20000, Grifola frondosa, SDS-PAGE
Grifola frondosa
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.4.24.20
Grifola frondosa
-
i.e. Maitake, var. S.F.Gray
-
Purification (Commentary)
EC Number
Commentary
Organism
3.4.24.20
-
Grifola frondosa
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.4.24.20
fruit body
-
Grifola frondosa
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.4.24.20
additional information
-
-
Grifola frondosa
Storage Stability
EC Number
Storage Stability
Organism
3.4.24.20
-40C, pH 7, at least a year
Grifola frondosa
3.4.24.20
4C, in various buffers, pH 5-10, about 20% loss of activity within 18 h
Grifola frondosa
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.4.24.20
azocasein + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Azocoll + H2O
general protease substrate
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Bovine serum albumin + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Congo red elastin + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Equine heart cytochrome c + H2O
cleaves-X-Lys bonds
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
additional information
specific cleavage of acyl-Lys bonds (-X-Lys-) in polypeptides
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
protamine sulfate + H2O
-
31047
Grifola frondosa
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
3.4.24.20
?
x * 20000, Grifola frondosa, SDS-PAGE
Grifola frondosa
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.4.24.20
37
-
assay at
Grifola frondosa
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
3.4.24.20
50
-
3 h, about 30% loss of activity at pH 9
Grifola frondosa
3.4.24.20
60
-
3 h, about 55% loss of activity at pH 9
Grifola frondosa
3.4.24.20
70
-
3 h, about 40% loss of activity with Mn2+ instead of zinc; 3 h, about 90% loss of activity at pH 9; and below, 3 h stable at pH 7.2
Grifola frondosa
3.4.24.20
80
-
3 h, 40% loss of activity at pH 7.2, 30 min, about 90% loss of activity with Mn2+ instead of zinc
Grifola frondosa
3.4.24.20
100
-
30 min, inactivation
Grifola frondosa
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.4.24.20
additional information
-
pI: 7.46
Grifola frondosa
3.4.24.20
9
-
congo red elastin as substrate
Grifola frondosa
3.4.24.20
9.5
-
azocoll or azocasein as substrate
Grifola frondosa
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.4.24.20
6
10.5
more than 50% of maximal activity at pH 6 and 10.5
Grifola frondosa
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.4.24.20
5
10
18 h, about 20% loss of activity at 4C
Grifola frondosa
3.4.24.20
7.2
-
3 h, stable at 70C and below, 40% loss of activity at 80C, inactivation within 30 min at 100C
Grifola frondosa
3.4.24.20
9
-
3 h, about 30%, 55% and 90% loss of activity at 50C, 60C and 70C, respectively
Grifola frondosa
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.4.24.20
1,10-phenanthroline
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
2,2'-bipyridine
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
2-mercaptoethanol
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
6-aminohexanoic acid
caseinolytic activity
Grifola frondosa
3.4.24.20
agmatine
caseinolytic activity
Grifola frondosa
3.4.24.20
antipain
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Ba2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Ca2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
Co2+
0.1 mM, caseinolytic activity
Grifola frondosa
3.4.24.20
Cu2+
1 mM, caseinolytic activity
Grifola frondosa
3.4.24.20
dithiothreitol
caseinolytic activity; strong
Grifola frondosa
3.4.24.20
DL-homoarginine
caseinolytic activity
Grifola frondosa
3.4.24.20
EDTA
fully restorable by Ca2+; fully restorable by Zn2+, Mn2+, Co2+; strong, caseinolytic activity
Grifola frondosa
3.4.24.20
Hg2+
strong, caseinolytic activity
Grifola frondosa
3.4.24.20
L-aminobutyric acid
caseinolytic activity
Grifola frondosa
3.4.24.20
L-arginine
caseinolytic activity
Grifola frondosa
3.4.24.20
L-lysine
caseinolytic activity
Grifola frondosa
3.4.24.20
L-Lysine ethylester
caseinolytic activity
Grifola frondosa
3.4.24.20
L-ornithine
caseinolytic activity
Grifola frondosa
3.4.24.20
Mg2+
1 mM, weak, caseinolytic activity
Grifola frondosa
3.4.24.20
additional information
no inhibition (of caseinolytic activity) by chymostatin, iodoacetate, leupeptin; PMSF; zinc acetate, elastatinal, Lys-Lys, 4 M urea
Grifola frondosa
3.4.24.20
p-chloromercuribenzoate
not
Grifola frondosa
3.4.24.20
phosphoramidon
weak, caseinolytic activity
Grifola frondosa
3.4.24.20
tosyl-Lys chloromethyl ketone
not
Grifola frondosa
3.4.24.20
tosyl-Phe chloromethyl ketone
weak, caseinolytic activity
Grifola frondosa
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.4.24.20
cytosol
-
Grifola frondosa
5829
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.4.24.20
Ca2+
activation, zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Co2+
activation, zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Mn2+
activation, 0.1 mM, native enzyme and zinc-depleted apoenzyme
Grifola frondosa
3.4.24.20
Zinc
atomic absorption spectroscopy; requirement, 1 atom per molecule of native enzyme; Zn2+ activates zinc-depleted apoenzyme
Grifola frondosa
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.4.24.20
20000
-
x * 20000, Grifola frondosa, SDS-PAGE
Grifola frondosa
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.4.24.20
-
Grifola frondosa
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.4.24.20
fruit body
-
Grifola frondosa
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.4.24.20
additional information
-
-
Grifola frondosa
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
3.4.24.20
-40C, pH 7, at least a year
Grifola frondosa
3.4.24.20
4C, in various buffers, pH 5-10, about 20% loss of activity within 18 h
Grifola frondosa
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.4.24.20
azocasein + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Azocoll + H2O
general protease substrate
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Bovine serum albumin + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Congo red elastin + H2O
-
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
Equine heart cytochrome c + H2O
cleaves-X-Lys bonds
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
additional information
specific cleavage of acyl-Lys bonds (-X-Lys-) in polypeptides
31047
Grifola frondosa
?
-
-
-
-
3.4.24.20
protamine sulfate + H2O
-
31047
Grifola frondosa
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.4.24.20
?
x * 20000, Grifola frondosa, SDS-PAGE
Grifola frondosa
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.4.24.20
37
-
assay at
Grifola frondosa
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
3.4.24.20
50
-
3 h, about 30% loss of activity at pH 9
Grifola frondosa
3.4.24.20
60
-
3 h, about 55% loss of activity at pH 9
Grifola frondosa
3.4.24.20
70
-
3 h, about 40% loss of activity with Mn2+ instead of zinc; 3 h, about 90% loss of activity at pH 9; and below, 3 h stable at pH 7.2
Grifola frondosa
3.4.24.20
80
-
3 h, 40% loss of activity at pH 7.2, 30 min, about 90% loss of activity with Mn2+ instead of zinc
Grifola frondosa
3.4.24.20
100
-
30 min, inactivation
Grifola frondosa
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.4.24.20
additional information
-
pI: 7.46
Grifola frondosa
3.4.24.20
9
-
congo red elastin as substrate
Grifola frondosa
3.4.24.20
9.5
-
azocoll or azocasein as substrate
Grifola frondosa
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.4.24.20
6
10.5
more than 50% of maximal activity at pH 6 and 10.5
Grifola frondosa
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.4.24.20
5
10
18 h, about 20% loss of activity at 4C
Grifola frondosa
3.4.24.20
7.2
-
3 h, stable at 70C and below, 40% loss of activity at 80C, inactivation within 30 min at 100C
Grifola frondosa
3.4.24.20
9
-
3 h, about 30%, 55% and 90% loss of activity at 50C, 60C and 70C, respectively
Grifola frondosa