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Literature summary extracted from

  • Burlini, N.; Magnani, P.; Villa, A.; Macchi, F.; Tortora, P.; Guerritore, A.
    A heat-stable serine proteinase from the extreme thermophilic archaebacterium Sulfolobus solfataricus (1992), Biochim. Biophys. Acta, 1122, 283-292.
    View publication on PubMed

General Stability

EC Number General Stability Organism
3.4.21.B52 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 Saccharolobus solfataricus
3.4.21.B52 the enzyme loses 54% of its activity after 60 min in buffer containing 2 mM dithiothreitol at 95°C Saccharolobus solfataricus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.21.B52 chymostatin 0.3 mg/ml, 94% inhibition Saccharolobus solfataricus
3.4.21.B52 iodoacetate 1 mM, 10% inhibition Saccharolobus solfataricus
3.4.21.B52 N-alpha-p-tosyl-L-lysine chloromethyl ketone 0.3 mM, 6% inhibition Saccharolobus solfataricus
3.4.21.B52 p-chloromercuribenzoate 0.6 mM, 6% inhibition Saccharolobus solfataricus
3.4.21.B52 phenylmethanesulfonyl fluoride 5 mM, 98% inhibition Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.B52 0.11
-
 benzyloxycarbonyl-Tyr-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 0.34
-
 benzyloxycarbonyl-Leu-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 0.72
-
 Suc-Gly-Gly-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 0.85
-
 Suc-Ala-Ala-Pro-Phe 4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 1.16
-
 Suc-Ala-Ala-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 1.34
-
 Suc-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.21.B52 intracellular
-
 Saccharolobus solfataricus 5622
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.21.B52 Co2+ activation effect up to 2 mM Saccharolobus solfataricus
3.4.21.B52 Mg2+ no effect by Mg2+, Ca2+ and Zn2+ over the concentration range 0.2-20 mM Saccharolobus solfataricus
3.4.21.B52 Mn2+ significantly stimulates, mixed type non-essential activator Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.21.B52 54000
-
 2 * 54000, SDS-PAGE Saccharolobus solfataricus
3.4.21.B52 118000
-
 gel filtration Saccharolobus solfataricus

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.4.21.B52 acetonitrile 50% acetonitrile, 91% inhibition after 30 min at 50°C Saccharolobus solfataricus
3.4.21.B52 Ethanol 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 Saccharolobus solfataricus
3.4.21.B52 Glycerol 20% glycerol, 35% inhibition after 30 min at 95°C Saccharolobus solfataricus
3.4.21.B52 Methanol 50% methanol, 66% loss of activity after 30 min at 50°C, 98% loss of activity after 30 min, at 95°C Saccharolobus solfataricus
3.4.21.B52 tetrahydrofuran 50% tetrahydrofuran, 77% inhibition after 30 min at 50°C, 96% inhibition after 30 min at 95°C Saccharolobus solfataricus
3.4.21.B52 urea enzyme can withstand 6 M urea at 50°C, 97% loss of activity after 30 min at 95°C Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.B52 Saccharolobus solfataricus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.B52
-
 Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.21.B52 28.9
-
 pH 7.0, 80°C Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.B52 benzyloxycarbonyl-Ala-Ala-Leu-4-nitroanilide + H2O 1% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide Saccharolobus solfataricus benzyloxycarbonyl-Ala-Ala-Leu + 4-nitroaniline
-
 ?
3.4.21.B52 benzyloxycarbonyl-Leu-4-nitroanilide + H2O 36% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide Saccharolobus solfataricus benzyloxycarbonyl-Leu + 4-nitroaniline
-
 ?
3.4.21.B52 benzyloxycarbonyl-Tyr-4-nitroanilide + H2O
-
 Saccharolobus solfataricus benzyloxycarbonyl-Tyr + 4-nitroaniline
-
 ?
3.4.21.B52 additional information 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 Saccharolobus solfataricus ?
-
 ?
3.4.21.B52 Phe-Ser-Lys-Phe-Ile-Ser-Phe-Asp + H2O no cleavage between Phe4 and Ile5 Saccharolobus solfataricus Phe-Ser-Lys-Phe-Ile + Ser-Phe-Asp
-
 ?
3.4.21.B52 Suc-Ala-Ala-Phe-4-nitroanilide + H2O 3% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide Saccharolobus solfataricus Suc-Ala-Ala-Phe + 4-nitroaniline
-
 ?
3.4.21.B52 Suc-Ala-Ala-Pro-Phe-4-nitroanilide + H2O 2% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide Saccharolobus solfataricus Suc-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?
3.4.21.B52 Suc-Gly-Gly-Phe-4-nitroanilide + H2O 6.5% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide Saccharolobus solfataricus Suc-Gly-Gly-Phe + 4-nitroaniline
-
 ?
3.4.21.B52 Suc-Phe-4-nitroanilide + H2O 18.5% of the activity with benzyloxycarbonyl-Tyr-p-nitroanilide Saccharolobus solfataricus Suc-Phe + 4-nitroaniline
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.21.B52 dimer 2 * 54000, SDS-PAGE Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.21.B52 50
-
 enzyme can withstand 6 M urea Saccharolobus solfataricus
3.4.21.B52 90
-
 completly stable up to Saccharolobus solfataricus
3.4.21.B52 92
-
 first-order thermal inactivation, half-life: 342 min Saccharolobus solfataricus
3.4.21.B52 101
-
 first-order thermal inactivation, half-life: 7 min Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.B52 1
-
 Suc-Ala-Ala-Pro-Phe 4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 2.2
-
 Suc-Ala-Ala-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 3.3
-
 Suc-Gly-Gly-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 12.1
-
 benzyloxycarbonyl-Leu-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 15.6
-
 Suc-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 23
-
 benzyloxycarbonyl-Tyr-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.21.B52 6.5 8
-
 Saccharolobus solfataricus
3.4.21.B52 7.5
-
 in absence of divalent cations Saccharolobus solfataricus

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.4.21.B52 Saccharolobus solfataricus isoelectric focusing
-
 5.6

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.21.B52 1.2
-
 Suc-Ala-Ala-Pro-Phe 4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 1.9
-
 Suc-Ala-Ala-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 4.6
-
 Suc-Gly-Gly-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 10.7
-
 Suc-Phe-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 35
-
 benzyloxycarbonyl-Leu-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus
3.4.21.B52 210
-
 benzyloxycarbonyl-Tyr-4-nitroanilide pH 7.0, 80°C Saccharolobus solfataricus