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

BRENDA support

Literature summary for 3.4.24.27 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
Search for more literature for 3.4.24.27

Activating Compound

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

Cloned(Commentary)

Cloned (Comment) Organism
gene npr, expression of wild-type and mutant enzymes in Escherichia coli K12 strain JM109 Bacillus thermoproteolyticus

Protein Variants

Protein Variants Comment Organism
Q128A site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
Q128E site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
Q128K site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
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
Q225D site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Bacillus thermoproteolyticus
Q225E site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
Q225K site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Bacillus thermoproteolyticus
Q225R site-directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.24
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225V Bacillus thermoproteolyticus
0.28
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128A Bacillus thermoproteolyticus
0.29
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225A Bacillus thermoproteolyticus
0.3
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutants Q128K and Q225R Bacillus thermoproteolyticus
0.33
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q128E Bacillus thermoproteolyticus
0.34
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225D Bacillus thermoproteolyticus
0.39
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225K Bacillus thermoproteolyticus
0.45
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant wild-type enzyme Bacillus thermoproteolyticus
0.63
-
 N-carbobenzoxy-L-Asp-L-Phe methyl ester pH 7.5, 25°C, recombinant mutant Q225E Bacillus thermoproteolyticus

Metals/Ions

Metals/Ions Comment Organism Structure
K+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ Bacillus thermoproteolyticus
Li+ activates, preference of monovalent cations in descending order: Na+, K+, Li+ Bacillus thermoproteolyticus
additional information mechanism of salt-induced activation Bacillus thermoproteolyticus
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
Zn2+ stereochemical relationships between Gln128, Glu143, Gln225, Asp226, His231 and active site Zn2+ of thermolysin, overview Bacillus thermoproteolyticus

Organism

Organism UniProt Comment Textmining
Bacillus thermoproteolyticus
-
 gene npr
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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)

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

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

pH Range

pH Minimum pH Maximum Comment Organism
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