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Literature summary extracted from

  • Yasukawa, K.; Inouye, K.
    Improving the activity and stability of thermolysin by site-directed mutagenesis (2007), Biochim. Biophys. Acta, 1774, 1281-1288.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.24.27 gene npr, DNA and amino acid sequence determination, expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109 Bacillus thermoproteolyticus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.24.27 G8C/N60C/S65P site-directed mutagenesis, the mutant shows a similar catalytic efficiency compared tot he wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 G8C/N60C/S65P/L144S site-directed mutagenesis, the mutant shows about 6fold increased catalytic effiency compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 L144S site-directed mutagenesis, the mutant shows about 10fold increased catalytic effiency compared to the wild-type enzyme Bacillus thermoproteolyticus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.24.27 additional information
-
 additional information kinetics and thermodynamics of wild-type and mutant enzymes in absence or presence of salt, overview Bacillus thermoproteolyticus
3.4.24.27 0.06
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant L144S Bacillus thermoproteolyticus
3.4.24.27 0.1
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P/L144S Bacillus thermoproteolyticus
3.4.24.27 0.18
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P Bacillus thermoproteolyticus
3.4.24.27 0.33
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant wild-type enzyme Bacillus thermoproteolyticus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.24.27 Ca2+ four ions per enzyme molecule required for stability Bacillus thermoproteolyticus
3.4.24.27 NaCl activates Bacillus thermoproteolyticus
3.4.24.27 Zn2+ one ion per enzyme molecule required for activity Bacillus thermoproteolyticus

Organism

EC Number Organism UniProt Comment Textmining
3.4.24.27 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-carbobenzoxy-L-Asp-L-Phe methyl ester + H2O
-
 Bacillus thermoproteolyticus ?
-
 ?
3.4.24.27 N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O i.e. FAGLA Bacillus thermoproteolyticus ?
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.24.27 25
-
 assay at Bacillus thermoproteolyticus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.24.27 3.3
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P Bacillus thermoproteolyticus
3.4.24.27 3.4
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 6.7
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant L144S Bacillus thermoproteolyticus
3.4.24.27 7
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P/L144S Bacillus thermoproteolyticus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.24.27 6.5 7
-
 Bacillus thermoproteolyticus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.24.27 additional information
-
 pH profile Bacillus thermoproteolyticus