Literature summary for 3.4.21.B52 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.

Search for more literature for 3.4.21.B52

General Stability

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

Inhibitors

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

KM Value [mM]

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

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
intracellular	-	Saccharolobus solfataricus	5622	-

Metals/Ions

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

Molecular Weight [Da]

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

Organic Solvent Stability

Organic Solvent	Comment	Organism
acetonitrile	50% acetonitrile, 91% inhibition after 30 min at 50°C	Saccharolobus solfataricus
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
Glycerol	20% glycerol, 35% inhibition after 30 min at 95°C	Saccharolobus solfataricus
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
tetrahydrofuran	50% tetrahydrofuran, 77% inhibition after 30 min at 50°C, 96% inhibition after 30 min at 95°C	Saccharolobus solfataricus
urea	enzyme can withstand 6 M urea at 50°C, 97% loss of activity after 30 min at 95°C	Saccharolobus solfataricus

Organism

Organism	UniProt	Comment	Textmining
Saccharolobus solfataricus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
benzyloxycarbonyl-Ala-Ala-Leu-4-nitroanilide + H2O	1% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide	Saccharolobus solfataricus	benzyloxycarbonyl-Ala-Ala-Leu + 4-nitroaniline	-	?
benzyloxycarbonyl-Leu-4-nitroanilide + H2O	36% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide	Saccharolobus solfataricus	benzyloxycarbonyl-Leu + 4-nitroaniline	-	?
benzyloxycarbonyl-Tyr-4-nitroanilide + H2O	-	Saccharolobus solfataricus	benzyloxycarbonyl-Tyr + 4-nitroaniline	-	?
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	?	-	?
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	-	?
Suc-Ala-Ala-Phe-4-nitroanilide + H2O	3% of the activity with benzyloxycarbonyl-Tyr-4-nitroanilide	Saccharolobus solfataricus	Suc-Ala-Ala-Phe + 4-nitroaniline	-	?
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	-	?
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	-	?
Suc-Phe-4-nitroanilide + H2O	18.5% of the activity with benzyloxycarbonyl-Tyr-p-nitroanilide	Saccharolobus solfataricus	Suc-Phe + 4-nitroaniline	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 54000, SDS-PAGE	Saccharolobus solfataricus

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	8	-	Saccharolobus solfataricus
7.5	-	in absence of divalent cations	Saccharolobus solfataricus

pI Value

Organism	Comment	pI Value Maximum	pI Value
Saccharolobus solfataricus	isoelectric focusing	-	5.6

kcat/KM [mM/s]

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

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Release 2023.1 (January 2023)