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Literature summary extracted from

  • Kusano, M.; Yasukawa, K.; Inouye, K.
    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.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
3.4.24.27 A113D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 A113E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 A113H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 A113K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 A113R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 D150A 131% residual activity with casein, 81% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 D150E 128% residual activity with casein, 228% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 D150K 51% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 D150R 44% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 D170A complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 D170E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 D170H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 D170K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 D170R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 F114A 28% residual activity with casein, 8% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 F114D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 F114E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 F114H 18% residual activity with casein, 20% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 F114K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 F114R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162A complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 G162R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 I168D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 I168E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 I168K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 I168R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 N227D 11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 N227E 36% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 N227K 29% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 N227R 55% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 S169A 112% residual activity with casein, 64% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 S169D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 S169E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 S169H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 S169K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 S169R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 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
3.4.24.27 S234D 5% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 S234E 4% residual activity with casein, 7% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 S234H 32% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 S234K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 S234R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 V230A 17% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 V230D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 V230E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 V230H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 V230K 3% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 V230R 6% residual activity with casein, 12% residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 W115A complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 W115D complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 W115E complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 W115H complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 W115K complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 W115R complete loss of activity Bacillus thermoproteolyticus
3.4.24.27 Y157A 11% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 Y157D 1% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 Y157E 13% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 Y157H 24% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 Y157K 7% residual activity with casein, no residual activity with substrate N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide Bacillus thermoproteolyticus
3.4.24.27 Y157R complete loss of activity Bacillus thermoproteolyticus

Organism

EC Number Organism UniProt Comment Textmining
3.4.24.27 Bacillus thermoproteolyticus
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.24.27 0.34
-
 mutant F114H, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.42
-
 mutant I168H, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.42
-
 mutant S234A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.55
-
 mutant N227A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.62
-
 mutant D150W, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.68
-
 mutant I168A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.84
-
 mutant D150H, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.89
-
 mutant D150A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.89
-
 mutant S169A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 0.93
-
 mutant D150E, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 1.08
-
 wild-type, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 1.14
-
 mutant F114A, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus
3.4.24.27 1.18
-
 mutant N227H, substrate casein, 25°C, pH 7.5 Bacillus thermoproteolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.24.27 casein + H2O
-
 Bacillus thermoproteolyticus ?
-
 ?
3.4.24.27 N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide + H2O
-
 Bacillus thermoproteolyticus ?
-
 ?