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Literature summary for 3.4.24.27 extracted from

  • Matsumiya, Y.; Murata, N.; Inouye, K.; Kubo, M.
    Further stabilization of Leu155 mutant thermolysins by mutation of an autodegradation site (2011), Appl. Biochem. Biotechnol., 166, 735-743.
    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 Bacillus subtilis by vector pTB51, the enzymes are secreted to the culture medium Bacillus sp. (in: Bacteria)

Protein Variants

Protein Variants Comment Organism
L155A site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
L155A/I156N site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
L155A/I156V site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
L155S site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is slightly enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
L155S/I156N site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)
L155S/I156V site-directed mutagenesis, the mutation abolishes the autodegradation activity, mutant thermostability at 80°C is enhanced compared to the wild-type enzyme Bacillus sp. (in: Bacteria)

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
12
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
12
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
13
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
13
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
13
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
14
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant wild-type enzyme, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
20
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A, pH 7.0, 37°C Bacillus sp. (in: Bacteria)

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability Bacillus sp. (in: Bacteria)
Zn2+ TLN contains one zinc ion and four calcium ions, and these ions contribute to enzymatic catalysis and structural stability Bacillus sp. (in: Bacteria)

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Bacillus sp. (in: Bacteria) autodegradation at position 154–155 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Bacillus sp. (in: Bacteria)
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification autodegradation at position 154-155 Bacillus sp. (in: Bacteria)

Purification (Commentary)

Purification (Comment) Organism
recombinant extracellular wild-type and mutant enzymes from Bacillus subtilis culture medium by ammonium sulfate fractionation, hydrophobic interaction chromatography, and gel filtration Bacillus sp. (in: Bacteria)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information autodegradation at position 154–155 Bacillus sp. (in: Bacteria) ?
-
 ?
N-benzyloxycarbonyl-Gly-L-Leu amide + H2O
-
 Bacillus sp. (in: Bacteria) N-benzyloxycarbonyl-Gly + L-Leu amide
-
 ?

Synonyms

Synonyms Comment Organism
TLN
-
 Bacillus sp. (in: Bacteria)

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Bacillus sp. (in: Bacteria)

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 half-life of wild-type enzyme is 18.3 min, half-lives of mutants are 25 min for L155A, 24 min for L155S, 60.8 min for L155A/I156V, 62.4 min for L155S/I156N, 93.3 for L155S/I156N, and 40 min for L155S/I156V Bacillus sp. (in: Bacteria)

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
13.58
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
26.67
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
48.33
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
51.67
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant wild-type enzyme, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
80
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
105
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
165
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S, pH 7.0, 37°C Bacillus sp. (in: Bacteria)

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Bacillus sp. (in: Bacteria)

General Information

General Information Comment Organism
additional information autodegradation at position 154–155 Bacillus sp. (in: Bacteria)

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.03
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
2
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
3.67
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant wild-type enzyme, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
4
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155A, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
4
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156N, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
12
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S/I156V, pH 7.0, 37°C Bacillus sp. (in: Bacteria)
12.67
-
 N-benzyloxycarbonyl-Gly-L-Leu amide recombinant mutant L155S, pH 7.0, 37°C Bacillus sp. (in: Bacteria)