Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Tatsumi, C.; Hashida, Y.; Yasukawa, K.; Inouye, K.
    Effects of site-directed mutagenesis of the surface residues Gln128 and Gln225 of thermolysin on its catalytic activity (2007), J. Biochem., 141, 835-842.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.4.24.27 additional information mechanism of salt-induced activation Bacillus thermoproteolyticus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.24.27 gene npr, 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 Q128A site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q128E site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q128K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225A site-directed mutagenesis, the mutant shows altered pKa value and stimulation of activity by NaCl and reduced activity with the negatively charged substrate N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester substrate compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225D site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225E site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225K site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225R site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 Q225V site-directed mutagenesis, the mutant shows altered pKa value and stimulation of activity by NaCl and reduced activity with the negatively charged substrate N-carbobenzoxy-L-aspartyl-L-phenylalanine methyl ester substrate 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 0.24
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225V Bacillus thermoproteolyticus
3.4.24.27 0.28
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128A Bacillus thermoproteolyticus
3.4.24.27 0.29
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225A Bacillus thermoproteolyticus
3.4.24.27 0.3
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutants Q128K and Q225R Bacillus thermoproteolyticus
3.4.24.27 0.33
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128E Bacillus thermoproteolyticus
3.4.24.27 0.34
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225D Bacillus thermoproteolyticus
3.4.24.27 0.39
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225K Bacillus thermoproteolyticus
3.4.24.27 0.45
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 0.63
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225E Bacillus thermoproteolyticus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.24.27 K+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ Bacillus thermoproteolyticus
3.4.24.27 Li+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ Bacillus thermoproteolyticus
3.4.24.27 additional information mechanism of salt-induced activation Bacillus thermoproteolyticus
3.4.24.27 Na+ activates, preference of monovalent cations in descending order: Na+, K+, Li+, the bell-shaped pH dependence profile of the FAGLA-hydrolyzing activity of thermolysin is shifted from pH 5.4 to pH 6.7 by the addition of 4 M NaCl Bacillus thermoproteolyticus
3.4.24.27 Zn2+ stereochemical relationships between Gln128, Glu143, Gln225, Asp226, His231 and active site Zn2+ of thermolysin, overview Bacillus thermoproteolyticus

Organism

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

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 additional information
-
catalytic efficiency of wild-type and mutant enzymes in absence or presence of 4 M NaCl, overview Bacillus thermoproteolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 1.3
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225A Bacillus thermoproteolyticus
3.4.24.27 1.6
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225V Bacillus thermoproteolyticus
3.4.24.27 3.3
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225D Bacillus thermoproteolyticus
3.4.24.27 3.6
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225R Bacillus thermoproteolyticus
3.4.24.27 3.7
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128K Bacillus thermoproteolyticus
3.4.24.27 3.8
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128A Bacillus thermoproteolyticus
3.4.24.27 4.3
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutants Q128E and Q225E Bacillus thermoproteolyticus
3.4.24.27 4.7
-
N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant wild-type enzyme and mutant Q225K Bacillus thermoproteolyticus

pH Optimum

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

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.24.27 additional information
-
bell-shaped pH-dependence with importance of surfacecharges of thermolysin, the bell-shaped pH dependence profile of the FAGLA-hydrolyzing activity of thermolysin is shifted from pH 5.4 to pH 6.7 by the addition of 4 M NaCl Bacillus thermoproteolyticus