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Literature summary extracted from

  • Yasukawa, K.; Kusano, M.; Inouye, K.
    A new method for the extracellular production of recombinant thermolysin by co-expressing the mature sequence and pro-sequence in Escherichia coli (2007), Protein Eng. Des. Sel., 20, 375-383.
    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 and secretion to the cell culture medium Bacillus thermoproteolyticus

Protein Variants

EC Number Protein Variants Comment Organism
3.4.24.27 E143A site-directed mutagenesis, E143A might exist as a complex with the propetide in the supernatant, inactive mutant, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 additional information a mutant thermolysin is affected by its autocatalytic digestion activity Bacillus thermoproteolyticus
3.4.24.27 N112A site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 N112D site-directed mutagenesis, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 N112E site-directed mutagenesis, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 N112H site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 N112K site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected Bacillus thermoproteolyticus
3.4.24.27 N112R site-directed mutagenesis, inactive mutant, the autocatalytic activity is affected Bacillus thermoproteolyticus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.24.27 TMP1
-
 Bacillus thermoproteolyticus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.24.27 extracellular
-
 Bacillus thermoproteolyticus
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.4.24.27 Ca2+ four ions per enzyme molecule required for stability Bacillus thermoproteolyticus
3.4.24.27 Zn2+ zinc metalloproteinase, one ion per enzyme molecule required for activity Bacillus thermoproteolyticus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.24.27 26000
-
 x * 22 852, propeptide of thermolysin, sequence calculation, x * 26000, mature enzyme, SDS-PAGE Bacillus thermoproteolyticus

Organism

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

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.4.24.27 proteolytic modification thermolysin is synthesized as inactive pre-proenzyme and receives autocatalytic cleavage of the peptide bond linking the pro- and mature sequences, overview Bacillus thermoproteolyticus

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.24.27 recombinant wild-type and mutant enzymes from Escherichia coli K12 strain JM109 by hydrophobic interaction and Gly-D-Phe affinity chromatography Bacillus thermoproteolyticus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.24.27 casein + H2O
-
 Bacillus thermoproteolyticus ?
-
 ?
3.4.24.27 additional information thermolysin is a thermostable neutral metalloproteinase and performs autocatalytic cleavage for pro-enzyme activation Bacillus thermoproteolyticus ?
-
 ?
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 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.24.27 ? x * 22 852, propeptide of thermolysin, sequence calculation, x * 26000, mature enzyme, SDS-PAGE 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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.24.27 6.5 7
-
 Bacillus thermoproteolyticus