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Literature summary for 3.4.24.27 extracted from
- Insights into the catalytic roles of the polypeptide regions in the active site of thermolysin and generation of the thermolysin variants with high activity and stability (2009), J. Biochem., 145, 103-113.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A113D | complete loss of activity | Bacillus thermoproteolyticus |
| A113E | complete loss of activity | Bacillus thermoproteolyticus |
| A113H | complete loss of activity | Bacillus thermoproteolyticus |
| A113K | complete loss of activity | Bacillus thermoproteolyticus |
| A113R | complete loss of activity | Bacillus thermoproteolyticus |
| D150A | 131% residual activity with casein, 81% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| D150E | 128% residual activity with casein, 228% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| D150H | 105% residual activity with casein, 37% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| D150K | 51% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| D150R | 44% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| D150W | 81% residual activity with casein, 60% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| D170A | complete loss of activity | Bacillus thermoproteolyticus |
| D170E | complete loss of activity | Bacillus thermoproteolyticus |
| D170H | complete loss of activity | Bacillus thermoproteolyticus |
| D170K | complete loss of activity | Bacillus thermoproteolyticus |
| D170R | complete loss of activity | Bacillus thermoproteolyticus |
| F114A | 28% residual activity with casein, 8% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| F114D | complete loss of activity | Bacillus thermoproteolyticus |
| F114E | complete loss of activity | Bacillus thermoproteolyticus |
| F114H | 18% residual activity with casein, 20% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| F114K | complete loss of activity | Bacillus thermoproteolyticus |
| F114R | complete loss of activity | Bacillus thermoproteolyticus |
| G162A | complete loss of activity | Bacillus thermoproteolyticus |
| G162D | complete loss of activity | Bacillus thermoproteolyticus |
| G162E | complete loss of activity | Bacillus thermoproteolyticus |
| G162H | complete loss of activity | Bacillus thermoproteolyticus |
| G162K | complete loss of activity | Bacillus thermoproteolyticus |
| G162R | complete loss of activity | Bacillus thermoproteolyticus |
| I168A | 69% residual activity with casein, 125% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| I168D | complete loss of activity | Bacillus thermoproteolyticus |
| I168E | complete loss of activity | Bacillus thermoproteolyticus |
| I168H | 13% residual activity with casein, 35% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| I168K | complete loss of activity | Bacillus thermoproteolyticus |
| I168R | complete loss of activity | Bacillus thermoproteolyticus |
| N227A | 72% residual activity with casein , 28% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| N227D | 11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| N227E | 36% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| N227H | 19% residual activity with casein, 19% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| N227K | 29% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| N227R | 55% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| S169A | 112% residual activity with casein, 64% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| S169D | complete loss of activity | Bacillus thermoproteolyticus |
| S169E | complete loss of activity | Bacillus thermoproteolyticus |
| S169H | complete loss of activity | Bacillus thermoproteolyticus |
| S169K | complete loss of activity | Bacillus thermoproteolyticus |
| S169R | complete loss of activity | Bacillus thermoproteolyticus |
| S234A | 88% residual activity with casein, 17% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide. Thermal inactivation at 80°C is greatly suppressed | Bacillus thermoproteolyticus |
| S234D | 5% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| S234E | 4% residual activity with casein, 7% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| S234H | 32% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| S234K | complete loss of activity | Bacillus thermoproteolyticus |
| S234R | complete loss of activity | Bacillus thermoproteolyticus |
| V230A | 17% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| V230D | complete loss of activity | Bacillus thermoproteolyticus |
| V230E | complete loss of activity | Bacillus thermoproteolyticus |
| V230H | complete loss of activity | Bacillus thermoproteolyticus |
| V230K | 3% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| V230R | 6% residual activity with casein, 12% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| W115A | complete loss of activity | Bacillus thermoproteolyticus |
| W115D | complete loss of activity | Bacillus thermoproteolyticus |
| W115E | complete loss of activity | Bacillus thermoproteolyticus |
| W115H | complete loss of activity | Bacillus thermoproteolyticus |
| W115K | complete loss of activity | Bacillus thermoproteolyticus |
| W115R | complete loss of activity | Bacillus thermoproteolyticus |
| Y157A | 11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| Y157D | 1% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| Y157E | 13% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| Y157H | 24% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| Y157K | 7% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide | Bacillus thermoproteolyticus |
| Y157R | complete loss of activity | Bacillus thermoproteolyticus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus thermoproteolyticus | - |
- |
- |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.34 | - |
mutant F114H, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.42 | - |
mutant I168H, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.42 | - |
mutant S234A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.55 | - |
mutant N227A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.62 | - |
mutant D150W, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.68 | - |
mutant I168A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.84 | - |
mutant D150H, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.89 | - |
mutant D150A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.89 | - |
mutant S169A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 0.93 | - |
mutant D150E, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 1.08 | - |
wild-type, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 1.14 | - |
mutant F114A, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
| 1.18 | - |
mutant N227H, substrate casein, 25°C, pH 7.5 | Bacillus thermoproteolyticus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| casein + H2O | - |
Bacillus thermoproteolyticus | ? | - |
? |  |
| N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O | - |
Bacillus thermoproteolyticus | ? | - |
? | |
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