EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
3.4.21.62 | additional information | unlike subtilisin, subtilisin-like serine protease does not require propeptide for folding | Thermococcus kodakarensis |
EC Number | Application | Comment | Organism |
---|---|---|---|
3.4.21.62 | biotechnology | subtilisin-like serine protease has a great advantage over other proteases in high resistance to heat, denaturants, detergents and chelating agents and therefore has great potential for application in biotechnology fields | Thermococcus kodakarensis |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.4.21.62 | the gene encoding the 1689 derivative without a putative N-terminal signal sequence, Pro-subtilisin-like serine protease (Met + Ala1-Gly640), ligated into pET25b vector, mutant Pro-S359A overexpressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells | Thermococcus kodakarensis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.4.21.62 | S359A | active site mutant. Upon overproduction, Pro-S359A accumulates in the cells in a soluble form | Thermococcus kodakarensis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.4.21.62 | EDTA | subtilisin exhibits little activity at 80°C in the presence of 10 mM. Subtilisin-like serine protease is fully active even in the presence of 10 mM EDTA | Thermococcus kodakarensis | |
3.4.21.62 | guanidine hydrochloride | at 2 M inhibits subtilisin-like serine protease by 35% and at 4 M almost completely, whereas it has no inhibitory effect on subtilisin | Thermococcus kodakarensis | |
3.4.21.62 | additional information | subtilisin-like serine protease is fully resistant to treatment with 2-5% SDS, 4-8 M urea, 10% Tween-20 or 10% Triton X-100. In gel assay, subtilisin-like serine protease is fully denatured prior to SDS-PAGE by trichloroacetic acid treatment, followed by boiling for 5 min in the presence of SDS | Thermococcus kodakarensis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.21.62 | 0.11 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 20°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis | |
3.4.21.62 | 0.41 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 80°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis | |
3.4.21.62 | 2.4 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 20°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis | |
3.4.21.62 | 7.9 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 80°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.4.21.62 | additional information | unlike subtilisin, subtilisin-like serine protease does not require Ca2+ for folding | Thermococcus kodakarensis |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 44000 | - |
Pro-subtilisin-like serine protease derivative with N- and/or C-terminal truncation, gel filtration | Thermococcus kodakarensis |
3.4.21.62 | 44200 | - |
subtilisin-like serine protease, sequence analysis | Thermococcus kodakarensis |
3.4.21.62 | 55000 | - |
Pro-subtilisin-like serine protease derivative with N- and/or C-terminal truncation, gel filtration | Thermococcus kodakarensis |
3.4.21.62 | 65000 | - |
Pro-subtilisin-like serine protease, gel filtration | Thermococcus kodakarensis |
3.4.21.62 | 68630 | - |
Pro-subtilisin-like serine protease, sequence analysis | Thermococcus kodakarensis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.4.21.62 | Thermococcus kodakarensis | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.4.21.62 | subtilisin-like serine protease and Pro-S359A, at 4°C, by sonication, centrifugation, ammonium sulfate precipitation and gel filtration | Thermococcus kodakarensis |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 510 | - |
subtilisin-like serine protease | Thermococcus kodakarensis |
3.4.21.62 | 3100 | - |
subtilisin | Thermococcus kodakarensis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.4.21.62 | azocasein + H2O | - |
Thermococcus kodakarensis | ? | - |
? | |
3.4.21.62 | additional information | autoprocessing of Pro-subtilisin-like serine protease | Thermococcus kodakarensis | ? | - |
? | |
3.4.21.62 | N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O | - |
Thermococcus kodakarensis | N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline | - |
? | |
3.4.21.62 | oxidized insulin chain B + H2O | is cleaved by subtilisin-like serine protease at multiple sites, preferably at the C-termini of the hydrophobic residues, such as Tyr, Phe, Leu, Val and Ala. This peptide is also cleaved by subtilisin at the C-termini of the hydrophobic residues, but at more specific sites | Thermococcus kodakarensis | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.4.21.62 | monomer | 1* 42000, gel filtration, subtilisin-like serine protease. 1 * 65000, SDS-PAGE, Pro-S359A | Thermococcus kodakarensis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.4.21.62 | subtilisin | - |
Thermococcus kodakarensis |
3.4.21.62 | subtilisin-like serine protease | - |
Thermococcus kodakarensis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 100 | - |
subtilisin-like serine protease: 100°C, subtilisin: 90°C | Thermococcus kodakarensis |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 80 | - |
subtilisin-like serine protease is stable at 80°C for at least 3. It loses activity at 90°C and 100°C with half-lives of more than 3 h and 100 min, respectively. Subtilisin loses its activity at 100°C with a half-life of 50 min | Thermococcus kodakarensis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.21.62 | 1.6 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 20°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis | |
3.4.21.62 | 14 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 20°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis | |
3.4.21.62 | 25 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 80°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis | |
3.4.21.62 | 440 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 80°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 7 | 11.5 | subtilisin-like serine protease: pH 7.0-11.5, subtilisin: pH 8.0 and 11.5 | Thermococcus kodakarensis |
EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|---|
3.4.21.62 | 2 | 12 | subtilisin-like serine protease is fully stable between pH 7-11, whereas it is not fully stable below pH 6 and above pH 12. It loses more than 85% of its activity below pH 3 and at pH 13. Subtilisin is fully stable between pH 2-12 | Thermococcus kodakarensis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.4.21.62 | physiological function | Pro-subtilisin-like serine protease consists of an N-terminal propeptide (Ala1-Ala113), a mature domain (subtilisin-like serine protease, Val114-Val539) and a C-terminal propeptide (Asp540-Gly640) | Thermococcus kodakarensis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.4.21.62 | 6 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 20°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis | |
3.4.21.62 | 15 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 20°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis | |
3.4.21.62 | 56 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin, at 80°C, in 50 mM Tris-HCl buffer, pH 8.0 | Thermococcus kodakarensis | |
3.4.21.62 | 62 | - |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide | subtilisin-like serine protease, at 80°C, in 50 mM Tris-HCl buffer, pH 7.5 | Thermococcus kodakarensis |