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Literature summary for 3.4.21.B57 extracted from
- A novel extracellular subtilisin-like protease from the hyperthermophile Aeropyrum pernix K1: biochemical properties, cloning, and expression (2003), Extremophiles, 7, 391-399.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli of pernisine, lacking the leader sequence a fusion protein with glutathione-S-transferase | Aeropyrum pernix |
General Stability
| General Stability | Organism |
|---|---|
| 1 mM CaCl2 stabilizes the enzyme for up to 4 h at 120°C | Aeropyrum pernix |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 4-hydroxymercuribenzoate | 10 mM, 80% inhibition | Aeropyrum pernix |  |
| Aprotinin | - |
Aeropyrum pernix | |
| EDTA | 1 mM, 90% inhibition | Aeropyrum pernix | |
| EGTA | 1 mM, 94% inhibition | Aeropyrum pernix | |
| additional information | TPCK and TLCK, respectively chymotrypsin and trypsin-like inhibitors do not affect the activity. 1,10-phenanthroline has no effect | Aeropyrum pernix | |
| PMSF | 1 mM, 90% inhibition | Aeropyrum pernix | |
| Soybean trypsin inhibitor | 1 mg/ml, complete inhibition | Aeropyrum pernix |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | - |
Aeropyrum pernix | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | 1 mM, enhances activity. An increase in CaCl2 concentration did not affect further the enzyme activity | Aeropyrum pernix | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 34000 | - |
1 * 34000 SDS-PAGE | Aeropyrum pernix |
| 35160 | - |
gel filtration | Aeropyrum pernix |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| 2-mercaptoethanol | 5%, reduces the activity by 80% | Aeropyrum pernix |
| dithiothreitol | 5%, reduces the activity | Aeropyrum pernix |
| guanidine-HCl | 4 mM, 14% residual activity | Aeropyrum pernix |
| SDS | the enzyme retains 20% proteolytic activity | Aeropyrum pernix |
| urea | 4 mM, 44% residual activity | Aeropyrum pernix |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Aeropyrum pernix | - |
- |
- |
| Aeropyrum pernix DSM 11879 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Aeropyrum pernix |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Bovine serum albumin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix | ? | - |
? | |
| Bovine serum albumin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix DSM 11879 | ? | - |
? | |
| casein + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix | ? | - |
? | |
| casein + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix DSM 11879 | ? | - |
? | |
| Hemoglobin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix | ? | - |
? | |
| Hemoglobin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix DSM 11879 | ? | - |
? | |
| additional information | pernisine has no activity on N-alpha-benzoyl-D-Arg-4-nitroanilide, N-alpha-benzoyl-D-Tyr-4-nitroanilide, and N-succinyl-Ala-Ala-Ala-4-nitroanilide used to detect trypsin, chymotrypsin, and elastase activity, respectively. Aminopeptidase activity is not detected | Aeropyrum pernix | ? | - |
? | |
| additional information | pernisine has no activity on N-alpha-benzoyl-D-Arg-4-nitroanilide, N-alpha-benzoyl-D-Tyr-4-nitroanilide, and N-succinyl-Ala-Ala-Ala-4-nitroanilide used to detect trypsin, chymotrypsin, and elastase activity, respectively. Aminopeptidase activity is not detected | Aeropyrum pernix DSM 11879 | ? | - |
? | |
| ovalbumin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix | ? | - |
? | |
| ovalbumin + H2O | among the proteins tested, casein shows the highest degree of susceptibility with 100% of hydrolysis, while in the case of hemoglobin, ovalbumin, and bovine serum albumin it was about 50% | Aeropyrum pernix DSM 11879 | ? | - |
? | |
| succinyl-Ala-Ala-Pro-Leu-4-nitroanilide + H2O | - |
Aeropyrum pernix | succinyl-Ala-Ala-Pro-Leu + 4-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | - |
Aeropyrum pernix | succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 34000 SDS-PAGE | Aeropyrum pernix |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| pernisine | - |
Aeropyrum pernix |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
in the presence of 1 mM CaCl2 | Aeropyrum pernix |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 60 | 120 | active in a broad range of temperature, 60°C: about 35% of maximal activity, 120°C: 80% of maximal activity | Aeropyrum pernix |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
4 h, no loss of activity, in absence of Ca2+ | Aeropyrum pernix |
| 100 | - |
half life: 60 min, in absence of Ca2+ | Aeropyrum pernix |
| 110 | - |
half life: 40 min, in absence of Ca2+ | Aeropyrum pernix |
| 120 | - |
half life: 30 min, in absence of Ca2+ | Aeropyrum pernix |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | 9 | - |
Aeropyrum pernix |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 12 | pH 6.0: about 50% of maximal activity, pH 12.0: about 40% of maximal activity | Aeropyrum pernix |
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