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Literature summary for 3.4.21.62 extracted from

  • Pulido, M.A.; Koga, Y.; Takano, K.; Kanaya, S.
    Directed evolution of Tk-subtilisin from a hyperthermophilic archaeon: identification of a single amino acid substitution responsible for low-temperature adaptation (2007), Protein Eng. Des. Sel., 20, 143-153.
    View publication on PubMed
Search for more literature for 3.4.21.62

Cloned(Commentary)

Cloned (Comment) Organism
Escherichia coli HB101 transformed with plasmid pBR1560 containing the wild-type prepro-subtilisin. G56S-propeptide and His-tagged G56S-propeptide overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL transformed with vector pET25b and in Escherichia coli BL21(DE3) with vector pET28a, respectively Thermococcus kodakarensis

Protein Variants

Protein Variants Comment Organism
E61G pro-subtilisin mutant, exhibits a halo-forming activity at 80, 70 and 60°C Thermococcus kodakarensis
E61K pro-subtilisin mutant, exhibits a halo-forming activity at 80, 70 and 60°C Thermococcus kodakarensis
G56S pro-subtilisin mutant, does not seriously affect the structure of pro-subtilisin. Is more effectively matured than pro-subtilisin at lower temperatures Thermococcus kodakarensis
G56S/T135S low-temperature adaptation of prepro-G56S/T135S-subtilisin Thermococcus kodakarensis

Inhibitors

Inhibitors Comment Organism Structure
additional information enzymatic activity of mature subtilisin is inhibited by G56S-propeptide and wild-type propeptide in a concentration-dependent manner Thermococcus kodakarensis

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+
-
 Thermococcus kodakarensis

Organism

Organism UniProt Comment Textmining
Thermococcus kodakarensis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
G56S-propeptide and His-tagged G56S-propeptide purified by gel filtration 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-p-nitroanilide + H2O
-
 Thermococcus kodakarensis N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline
-
 ?

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 80 G56S-propeptide is almost completely degraded within 5–10 min at 80°C, 15–30 min at 60°C and 3 h at 40°C, but is not completely degraded upon incubation for 3 h at 20°C. Pro-subtilisin is rapidly autoprocessed into propeptide and mature subtilisin within 1 min even at 20°C. However, the subsequent degradation of propeptide proceeds much more slowly than that of G56S-propeptide. Propeptide is almost completely degraded only when it is incubated at 80°C for 30 min. It is not completely degraded for up to 3 h at 20, 40 and 60°C Thermococcus kodakarensis