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Information on EC 3.4.21.B52 - thermostable serine endopeptidase (Sulfolobus) for references in articles please use BRENDA:EC3.4.21.B52
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EC Tree
The expected taxonomic range for this enzyme is: Saccharolobus solfataricus
Reaction Schemes
The best substrate is benzyloxycarbonyl-Tyr-4-nitroanilide. Cleaves Phe-Ser-Lys-Phe-Ile-/-Ser-Phe-Asp. No activity benzyloxycarbonyl-Ile-4-nitroanilide, aminoacyl-4-nitroanilides or benzyloxycarbonyl-amino acids, oxidized insulin chain A and B, bovine pancreas ribonuclease A, bovine serum albumin or ovalbumin.
Synonyms
proteinase i,
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proteinase I
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The best substrate is benzyloxycarbonyl-Tyr-4-nitroanilide. Cleaves Phe-Ser-Lys-Phe-Ile-/-Ser-Phe-Asp. No activity benzyloxycarbonyl-Ile-4-nitroanilide, aminoacyl-4-nitroanilides or benzyloxycarbonyl-amino acids, oxidized insulin chain A and B, bovine pancreas ribonuclease A, bovine serum albumin or ovalbumin.
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benzyloxycarbonyl-Ala-Ala-Leu-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Leu + 4-nitroaniline
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1% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide
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?
benzyloxycarbonyl-Leu-4-nitroanilide + H2O
benzyloxycarbonyl-Leu + 4-nitroaniline
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36% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide
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?
benzyloxycarbonyl-Tyr-4-nitroanilide + H2O
benzyloxycarbonyl-Tyr + 4-nitroaniline
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?
Phe-Ser-Lys-Phe-Ile-Ser-Phe-Asp + H2O
Phe-Ser-Lys-Phe-Ile + Ser-Phe-Asp
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no cleavage between Phe4 and Ile5
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Suc-Ala-Ala-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Phe + 4-nitroaniline
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3% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide
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?
Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
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2% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide
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?
Suc-Gly-Gly-Phe-4-nitroanilide + H2O
Suc-Gly-Gly-Phe + 4-nitroaniline
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6.5% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide
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?
Suc-Phe-4-nitroanilide + H2O
Suc-Phe + 4-nitroaniline
additional information
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narrow substrate specificity. No activity with: aminoacyl-4-nitroanilides or benzyloxycarbonyl-amino acids, oxidized insulin chain A and B, bovine pancreas ribonuclease A, bovine serum albumin, ovalbumin. No activity with benzyloxycarbonyl-Ile-4-nitroanilide
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Suc-Phe-4-nitroanilide + H2O
Suc-Phe + 4-nitroaniline
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?
Suc-Phe-4-nitroanilide + H2O
Suc-Phe + 4-nitroaniline
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18.5% of the activity with benzyloxycarbonyl-Tyr-p-nitroanilide
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?
Suc-Phe-4-nitroanilide + H2O
Suc-Phe + 4-nitroaniline
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Co2+
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activation effect up to 2 mM
Mg2+
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no effect by Mg2+, Ca2+ and Zn2+ over the concentration range 0.2-20 mM
Mn2+
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significantly stimulates, mixed type non-essential activator
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iodoacetate
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1 mM, 10% inhibition
N-alpha-p-tosyl-L-lysine chloromethyl ketone
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0.3 mM, 6% inhibition
N-alpha-p-tosyl-L-phenylalanine chloromethyl ketone
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0.5 mM, complete inhibition
p-chloromercuribenzenesulfonate
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10 mM, 13% inhibition
p-chloromercuribenzoate
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0.6 mM, 6% inhibition
phenylmethanesulfonyl fluoride
chymostatin
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0.25 mg/ml, complete inhibition
chymostatin
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0.3 mg/ml, 94% inhibition
phenylmethanesulfonyl fluoride
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1 mM, 93% inhibition
phenylmethanesulfonyl fluoride
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5 mM, 98% inhibition
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0.34
benzyloxycarbonyl-Leu-4-nitroanilide
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pH 7.0, 80°C
0.11
benzyloxycarbonyl-Tyr-4-nitroanilide
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pH 7.0, 80°C
1.16
Suc-Ala-Ala-Phe-4-nitroanilide
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pH 7.0, 80°C
0.85
Suc-Ala-Ala-Pro-Phe 4-nitroanilide
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pH 7.0, 80°C
0.72
Suc-Gly-Gly-Phe-4-nitroanilide
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pH 7.0, 80°C
1.34
Suc-Phe-4-nitroanilide
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pH 7.0, 80°C
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12.1
benzyloxycarbonyl-Leu-4-nitroanilide
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pH 7.0, 80°C
23
benzyloxycarbonyl-Tyr-4-nitroanilide
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pH 7.0, 80°C
2.2
Suc-Ala-Ala-Phe-4-nitroanilide
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pH 7.0, 80°C
1
Suc-Ala-Ala-Pro-Phe 4-nitroanilide
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pH 7.0, 80°C
3.3
Suc-Gly-Gly-Phe-4-nitroanilide
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pH 7.0, 80°C
15.6
Suc-Phe-4-nitroanilide
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pH 7.0, 80°C
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35
benzyloxycarbonyl-Leu-4-nitroanilide
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pH 7.0, 80°C
210
benzyloxycarbonyl-Tyr-4-nitroanilide
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pH 7.0, 80°C
1.9
Suc-Ala-Ala-Phe-4-nitroanilide
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pH 7.0, 80°C
1.2
Suc-Ala-Ala-Pro-Phe 4-nitroanilide
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pH 7.0, 80°C
4.6
Suc-Gly-Gly-Phe-4-nitroanilide
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pH 7.0, 80°C
10.7
Suc-Phe-4-nitroanilide
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pH 7.0, 80°C
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7.5
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in absence of divalent cations
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5.6
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isoelectric focusing
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brenda
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brenda
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brenda
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brenda
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54000
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2 * 54000, SDS-PAGE
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dimer
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2 * 54000, SDS-PAGE
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50
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enzyme can withstand 6 M urea
92
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first-order thermal inactivation, half-life: 342 min
101
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first-order thermal inactivation, half-life: 7 min
90
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15 min, stable
90
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completly stable up to
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the enzyme is stable for 60 min in buffer containing 1 M NaCl at 50°C, it loses 96% of its activity after 60 min in 1 mM NaCl at 95°C
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the enzyme loses 54% of its activity after 60 min in buffer containing 2 mM dithiothreitol at 95°C
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acetonitrile
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50% acetonitrile, 91% inhibition after 30 min at 50°C
Ethanol
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enzyme can withstand 50% ethanol at 50°C, 98% loss of activity after 30 min at 95°C. 45% loss of activity after 30 min, in 99% ethanol
Glycerol
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20% glycerol, 35% inhibition after 30 min at 95°C
Methanol
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50% methanol, 66% loss of activity after 30 min at 50°C, 98% loss of activity after 30 min, at 95°C
tetrahydrofuran
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50% tetrahydrofuran, 77% inhibition after 30 min at 50°C, 96% inhibition after 30 min at 95°C
urea
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enzyme can withstand 6 M urea at 50°C, 97% loss of activity after 30 min at 95°C
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Fusi, P.; Villa, M.; Burlini, N.; Tortora, P.; Guerritore, A.
Intracellular proteases from the extremely thermophilic archaebacterium Sulfolobus solfataricus
Experientia
47
1057-1060
1991
Saccharolobus solfataricus, Saccharolobus solfataricus MT-4 / DSM 5833
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brenda
Burlini, N.; Magnani, P.; Villa, A.; Macchi, F.; Tortora, P.; Guerritore, A.
A heat-stable serine proteinase from the extreme thermophilic archaebacterium Sulfolobus solfataricus
Biochim. Biophys. Acta
1122
283-292
1992
Saccharolobus solfataricus
brenda
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3.4.21.B52 thermostable serine endopeptidase (Sulfolobus)
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