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Literature summary for 3.4.21.B48 extracted from
- Kumamolisin (2004), Handbook of proteolytic enzymes (Barrett, A. J. , Rawlings, N. D. , Woessner, J. F. , eds. ) Academic Press, 2nd Ed., 1889-1891.
No PubMed abstract available
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli strain JM109 | Bacillus novosp. |
| expression of wild-type and mutant enzymes in Escherichia coli strain JM109 | Weizmannia coagulans |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| enzyme in complex with inhibitors Ac-Ile-Ala-Phe-CHO and Ac-Ile-Pro-Phe-CHO, X-ray diffraction structure analysis at 1.4 A resolution | Weizmannia coagulans |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D164A | site-directed mutagenesis, no autoactivation of the proform, inactive mutant | Weizmannia coagulans |
| D316A | site-directed mutagenesis, autoactivation similar to the wild-type enzyme, 3% of wild-type enzyme activity | Weizmannia coagulans |
| D82A | site-directed mutagenesis, no autoactivation of the proform, inactive mutant | Weizmannia coagulans |
| E78A | site-directed mutagenesis, no autoactivation of the proform, inactive mutant | Weizmannia coagulans |
| S278A | site-directed mutagenesis, no autoactivation of the proform, inactive mutant | Weizmannia coagulans |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ac-Ile-Ala-Phe-CHO | synthetic aldehyde inhibitor, binds covalently to the active site, mechanism, overview | Weizmannia coagulans |  |
| Ac-Ile-Pro-Phe-CHO | synthetic aldehyde inhibitor, binds covalently to the active site, mechanism, overview | Weizmannia coagulans | |
| additional information | no inhibition by pepstatin, tyrostatin, DAN or EPNP | Weizmannia coagulans |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.041 | - |
Lys-Pro-Ile-Pro-Phe-(4-nitrophenyl)-Arg-Leu | pH 3.5, 60°C | Weizmannia coagulans | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 41000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Weizmannia coagulans |
| 41000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Bacillus novosp. |
| 43000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Weizmannia coagulans |
| 43000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Bacillus novosp. |
| 45000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Weizmannia coagulans |
| 45000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Bacillus novosp. |
| 64000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Weizmannia coagulans |
| 64000 | - |
x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Bacillus novosp. |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Bacillus novosp. | - |
- |
- |
| Bacillus novosp. MN-32 | - |
- |
- |
| Weizmannia coagulans | - |
- |
- |
| Weizmannia coagulans J-4 | - |
- |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | the enzyme needs to cleaved in to the mature enzyme and the N-terminal prepropeptide, autocatalytic cleavage | Weizmannia coagulans |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| specifically hydrolyzes Leu15-Tyr16 peptide bond in oxidized insulin B chain, additional cleavage at Phe25-Tyr26 at a considerably lower rate. Good cleavage of the Phe-I-(p-nitrophenylalanine) bond in synthetic peptides. The enzyme preferentially hydrolyzes peptides having an Ala or Pro residue at P2 position and prefers such charged amino acid residues as Glu or Arg at the P2' position. No cleavage: Asp-Pro-Ala-Lys-Phe-(p-nitrophenylalanine)-Arg-Leu | the catalytic triad is formed by Glu78, Asp82, and Ser278 | Weizmannia coagulans |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| additional information | high enzyme production conditions for large scale purification | Bacillus novosp. | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 6600 | - |
purified recombinant wild-type enzyme | Weizmannia coagulans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| Lys-Pro-Ile-Pro-Phe-(4-nitrophenyl)-Arg-Leu + H2O | - |
Weizmannia coagulans | Lys-Pro-Ile-Pro-Phe + (4-nitrophenyl)-Arg-Leu | - |
? | |
| Lys-Pro-Ile-Pro-Phe-(4-nitrophenyl)-Arg-Leu + H2O | - |
Weizmannia coagulans J-4 | Lys-Pro-Ile-Pro-Phe + (4-nitrophenyl)-Arg-Leu | - |
? | |
| additional information | substrate specificity, overview, the enzyme prefers charged amino acid residues at P2' position | Weizmannia coagulans | ? | - |
? | |
| additional information | substrate specificity, overview, the enzyme prefers charged amino acid residues at P2' position | Weizmannia coagulans J-4 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Weizmannia coagulans |
| ? | x * 41000, native wild-type enzyme, x * 43000, active recombinant wild-type enzyme, SDS-PAGE, x * 45000, inactive, processed recombinant wild-type enzyme, SDS-PAGE, x * 64000, inactive recombinant precursor enzyme, SDS-PAGE | Bacillus novosp. |
| More | the enzyme consists of an N-terminal prepropeptide and the mature enzyme part | Weizmannia coagulans |
| More | the enzyme consists of an N-terminal prepropeptide and the mature enzyme part | Bacillus novosp. |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| J-4 serine-carboxyl proteinase | - |
Weizmannia coagulans |
| KSCP | - |
Weizmannia coagulans |
| KSCP | - |
Bacillus novosp. |
| kumamolisin serine-carboxyl proteinase | - |
Weizmannia coagulans |
| kumamolisin serine-carboxyl proteinase | - |
Bacillus novosp. |
| More | the enzyme belongs to peptidase family 53 | Weizmannia coagulans |
| More | the enzyme belongs to peptidase family 53 | Bacillus novosp. |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 70 | - |
- |
Weizmannia coagulans |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 176 | - |
Lys-Pro-Ile-Pro-Phe-(4-nitrophenyl)-Arg-Leu | pH 3.5, 60°C | Weizmannia coagulans | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 3 | - |
- |
Weizmannia coagulans |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0007 | - |
Ac-Ile-Pro-Phe-CHO | pH 3.5, 22.4°C | Weizmannia coagulans | |
| 0.0009 | - |
Ac-Ile-Ala-Phe-CHO | pH 3.5, 22.4°C | Weizmannia coagulans | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Weizmannia coagulans | - |
- |
3.5 |
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