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Literature summary for 3.4.24.27 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)

Cloned (Comment) Organism
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

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

KM Value [mM]

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

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Bacillus thermoproteolyticus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
casein + H2O
-
Bacillus thermoproteolyticus ?
-
?
N-carbobenzoxy-L-Asp-L-Phe methyl ester + H2O
-
Bacillus thermoproteolyticus ?
-
?
N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide + H2O i.e. FAGLA Bacillus thermoproteolyticus ?
-
?

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5 7
-
Bacillus thermoproteolyticus

pH Range

pH Minimum pH Maximum Comment Organism
additional information
-
pH profile Bacillus thermoproteolyticus