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Literature summary extracted from

  • Takita, T.; Aono, T.; Sakurama, H.; Itoh, T.; Wada, T.; Minoda, M.; Yasukawa, K.; Inouye, K.
    Effects of introducing negative charges into the molecular surface of thermolysin by site-directed mutagenesis on its activity and stability (2008), Biochim. Biophys. Acta, 1784, 481-488.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.24.27 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 S198D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 S218D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 S254D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
3.4.24.27 S25D site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl Bacillus thermoproteolyticus
3.4.24.27 S53D site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl, increased thermostability in presence of 10 mM CaCl2 Bacillus thermoproteolyticus
3.4.24.27 S65D site directed mutagenesis, the catalytic activity is of the mutant enzyme is similar to the wild-type in absence of NaCl, but increased in presence of 4 M NaCl, increased thermostability in presence of 10 mM CaCl2 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 thermodynamics of wild-type and mutant enzymes Bacillus thermoproteolyticus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.24.27 Ca2+ required for stability Bacillus thermoproteolyticus
3.4.24.27 NaCl activates the mutant enzymes at 4 M to 17-19fold of wild-type enzyme activity, overview Bacillus thermoproteolyticus
3.4.24.27 Zn2+ required for activity Bacillus thermoproteolyticus

Organism

EC Number Organism UniProt Comment Textmining
3.4.24.27 Bacillus thermoproteolyticus P00800
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.24.27 recombinant wild-type and mutant enzymes from Escherichia coli K12 strain JM109 to homogeneity by Gly-D-Phe heat treatment at 60°C for 20 min, affinity chromatography and gel filtration Bacillus thermoproteolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.24.27 casein + H2O from bovine milk 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

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.24.27 85
-
 30 min, 51% remaining activity of recombinaqnt wild-type enzyme, 35-78% remaining activity of mutant enzymes Bacillus thermoproteolyticus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.24.27 7.5
-
 assay at Bacillus thermoproteolyticus