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Literature summary for 3.4.21.B57 extracted from

  • Pulido, M.; Saito, K.; Tanaka, S.; Koga, Y.; Morikawa, M.; Takano, K.; Kanaya, S.
    Ca2+-dependent maturation of subtilisin from a hyperthermophilic archaeon, Thermococcus kodakaraensis: the propeptide is a potent inhibitor of the mature domain but is not required for its folding (2006), Appl. Environ. Microbiol., 72, 4154-4162.
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
the propeptide and mature domain are independently overproduced in Escherichia coli Thermococcus kodakarensis

Inhibitors

Inhibitors Comment Organism Structure
Thermococcus kodakaraensis subtilisin propeptide potent noncompetitive inhibitor of the mature domain Thermococcus kodakarensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide 20°C, pH 9.5 Thermococcus kodakarensis
8
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide 80°C, pH 9.5, the Km value of 8.0 mM represents the apparent value, because the solubility of the substrate at pH 9.5 is too low to increase its concentration beyond 5 mM Thermococcus kodakarensis

Organism

Organism UniProt Comment Textmining
Thermococcus kodakarensis P58502 sequence including singnal peptide (amino acid 1-24) and propeptide (amino acid 25-106)
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification pro-subtilisin is inactive in the absence of Ca2+ but is activated upon autoprocessing and degradation of propeptide in the presence of Ca2+ at 80°C. This maturation process is completed within 30 min at 80°C but is bound at an intermediate stage, in which the propeptide is autoprocessed from the mature domain (mat-subtilisin) but forms an inactive complex with mat-subtilisin*, at lower temperatures. At 80°C, approximately 30% of the pro-subtilisin is autoprocessed into propeptide and mat-subtilisin, and the other 70% is completely degraded to small fragments. mat-Subtilisin is inactive in the absence of Ca2+ but is activated upon incubation with Ca2+ at 80°C Thermococcus kodakarensis

Purification (Commentary)

Purification (Comment) Organism
-
 Thermococcus kodakarensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
13
-
 20°C, pH 9.5 Thermococcus kodakarensis
140
-
 80°C, pH 9.5 Thermococcus kodakarensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
azocasein + H2O
-
 Thermococcus kodakarensis ?
-
 ?
N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O
-
 Thermococcus kodakarensis N-succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
-
 Thermococcus kodakarensis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 stable for at least 3 h Thermococcus kodakarensis
90
-
 the enzyme loses half of its activity in 9 h Thermococcus kodakarensis
100
-
 the enzyme loses half of its activity in 50 min Thermococcus kodakarensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
26
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide 20°C, pH 9.5 Thermococcus kodakarensis
290
-
 N-succinyl-Ala-Ala-Pro-Phe-4-nitroanilide 80°C, pH 9.5 Thermococcus kodakarensis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000025
-
 Thermococcus kodakaraensis subtilisin propeptide 20°C, pH 9.5 Thermococcus kodakarensis