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Literature summary for 3.4.24.27 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
Search for more literature for 3.4.24.27

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
S198D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
S218D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
S254D site directed mutagenesis, the mutant shows similar activity compared to the wild-type enzyme Bacillus thermoproteolyticus
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
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
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thermodynamics of wild-type and mutant enzymes Bacillus thermoproteolyticus

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Bacillus thermoproteolyticus P00800
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

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

Temperature Stability [°C]

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

pH Optimum

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