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Literature summary for 3.4.21.62 extracted from
- Purification and characterization of Ak.1 protease, a thermostable subtilisin with a disulphide bond in the substrate-binding cleft (2000), Biochem. J., 350, 321-328.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| AkP gene cloned and expressed in Escherichia coli PB5517 | Bacillus sp. (in: Bacteria) |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.07 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) |  |
| 0.15 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 0.28 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 0.3 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 0.32 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 0.33 | - |
Suc-Ala-Ala-Pro-Ala-p-nitroanilide | pH 7.5, 5°C or 40°C | Bacillus sp. (in: Bacteria) | |
| 0.44 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 0.58 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 0.63 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 0.68 | - |
Suc-Ala-Ala-Pro-Leu-p-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 0.92 | - |
Suc-Ala-Ala-Pro-Ala-p-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
| 1 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
| 1.01 | - |
Suc-Ala-Ala-Pro-Leu-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 2.25 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
| 3.37 | - |
Suc-Ala-Ala-Pro-Leu-p-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
| 4 | - |
Suc-Ala-Ala-p-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 6.67 | - |
Suc-Ala-Ala-p-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 8.3 | - |
benzyloxycarbonyl-Gly-4-nitroanilide | pH 7.5, 20°C, in presence and absence of DTT | Bacillus sp. (in: Bacteria) | |
| 17 | - |
Suc-Ala-Ala-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | bound calcium ions play a key role in protecting against autolysis and thermal denaturation | Bacillus sp. (in: Bacteria) | |
| Cd2+ | stabilizes | Bacillus sp. (in: Bacteria) | |
| Sr2+ | increases thermostability, but to a significantly lower degree than Ca2+ | Bacillus sp. (in: Bacteria) | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 36000 | - |
SDS-PAGE | Bacillus sp. (in: Bacteria) |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus sp. (in: Bacteria) | - |
strain Ak.1 | - |
| Bacillus sp. (in: Bacteria) Ak1 | - |
strain Ak.1 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme | Bacillus sp. (in: Bacteria) |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
specific activity of purified protease 174 units/mg, 1 unit = DELTA A420 of 1.0 per min at 75°C with 0.2% azocasein | Bacillus sp. (in: Bacteria) |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| benzyloxycarbonyl-Gly-4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | benzyloxycarbonyl-Gly + 4-nitroaniline | - |
? | |
| additional information | little or no reaction with Glu-Phe-p-nitroanilide, benzyloxycarbonyl-Phe-p-nitroanilide, Suc-Phe-p-nitroanilide, acetyl-Phe-p-nitroanilide, Phe-p-nitroanilide, Gly-p-nitroanilide and Ala-p-nitroanilide at 75°C, and Val-Leu-Lys-p-nitroanilide, benzoyl-Arg-p-nitroanilide, Suc-Ala-Ala-p-nitroanilide, benzoyl-Tyr-p-nitroanilide, benzyloxycarbonyl-Lys-Arg-p-nitroanilide, and benzyloxycarbonyl-Arg-p-nitroanilide at 40°C | Bacillus sp. (in: Bacteria) | ? | - |
? | |
| additional information | little or no reaction with Glu-Phe-p-nitroanilide, benzyloxycarbonyl-Phe-p-nitroanilide, Suc-Phe-p-nitroanilide, acetyl-Phe-p-nitroanilide, Phe-p-nitroanilide, Gly-p-nitroanilide and Ala-p-nitroanilide at 75°C, and Val-Leu-Lys-p-nitroanilide, benzoyl-Arg-p-nitroanilide, Suc-Ala-Ala-p-nitroanilide, benzoyl-Tyr-p-nitroanilide, benzyloxycarbonyl-Lys-Arg-p-nitroanilide, and benzyloxycarbonyl-Arg-p-nitroanilide at 40°C | Bacillus sp. (in: Bacteria) Ak1 | ? | - |
? | |
| N-succinyl-L-Ala-L-Ala-L-Pro-L-Leu-4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | ? | - |
? | |
| succinyl-AAPA-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | ? | - |
? | |
| succinyl-AAPF-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-AAPF + p-nitroaniline | - |
? | |
| succinyl-AAPF-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) Ak1 | succinyl-AAPF + p-nitroaniline | - |
? | |
| Succinyl-Ala-Ala 4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | ? | - |
? | |
| Succinyl-Ala-Ala 4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) Ak1 | ? | - |
? | |
| Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | ? | - |
? | |
| Succinyl-Ala-Ala-Ala 4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) Ak1 | ? | - |
? | |
| succinyl-Ala-Ala-Ala-Ala-Ala-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-AAAAA + p-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Ala-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-AAA + p-nitroaniline | - |
? | |
| succinyl-Ala-Ala-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-Ala-Ala + p-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Pro-Ala-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-AAPA + p-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-AAPL + p-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? | |
| succinyl-Ala-Ala-Pro-Phe-4-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) Ak1 | succinyl-Ala-Ala-Pro-Phe + 4-nitroaniline | - |
? | |
| succinyl-Phe-Ala-Ala-Phe-p-nitroanilide + H2O | - |
Bacillus sp. (in: Bacteria) | succinyl-Phe-Ala-Ala-Phe + p-nitroaniline | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Ak.1 protease | - |
Bacillus sp. (in: Bacteria) |
| thermostable subtilisin | - |
Bacillus sp. (in: Bacteria) |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 93 | - |
Suc-Ala-Ala-p-nitroanilide | pH 7.5, 20°C, in presence and absence of DTT | Bacillus sp. (in: Bacteria) | |
| 148 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 222 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 222 | - |
Suc-Ala-Ala-Pro-Ala-p-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 389 | - |
Suc-Ala-Ala-Pro-Leu-p-nitroanilide | pH 7.5, 5°C | Bacillus sp. (in: Bacteria) | |
| 750 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 1200 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 1900 | - |
benzyloxycarbonyl-Gly-4-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 2500 | - |
benzyloxycarbonyl-Gly-4-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 2900 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 20°C, DTT | Bacillus sp. (in: Bacteria) | |
| 3700 | - |
Suc-Ala-Ala-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 8800 | - |
Suc-Ala-Ala-Pro-Ala-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 10000 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 20°C | Bacillus sp. (in: Bacteria) | |
| 22000 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 97000 | - |
Suc-Ala-Ala-Pro-Leu-p-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 97000 | - |
Suc-Ala-Ala-Ala-p-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
| 130000 | - |
succinyl-Ala-Ala-Pro-Phe-4-nitroanilide | pH 7.5, 40°C | Bacillus sp. (in: Bacteria) | |
| 130000 | - |
Suc-Ala-Ala-Pro-Ala-p-nitroanilide | pH 7.5, 75°C | Bacillus sp. (in: Bacteria) | |
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