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Literature summary for 3.4.21.62 extracted from
- Collagenolytic subtilisin-like protease from the deep-sea bacterium Alkalimonas collagenimarina AC40T (2010), Appl. Microbiol. Biotechnol., 86, 589-598.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| vectors pRSET-acpII, pRSET-acpII-tr (truncated AcpII Met1-Arg675), pRSET-acpII-C (catalytic domain of AcpII) and pRSET-acpII-C-DELTAPA (AcpII-C with the protease-associated domain deleted) expressed in Escherichia coli BL21(DE3) pLysS | Alkalimonas collagenimarina |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | 5 mM partially inhibits by 35%, in 100 mM Tris-HCl buffer, pH 7.0 at 15°C for 20 min | Alkalimonas collagenimarina |  |
| phenylmethylsulfonyl fluoride | 1 mM completely inhibits, in 100 mM Tris-HCl buffer, pH 7.0 at 15°C for 20 min | Alkalimonas collagenimarina | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | enzyme activity and/or stability depends on the presence of divalent cations, probably Ca2+ ions, near the surface of the enzyme | Alkalimonas collagenimarina | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 55000 | - |
x * 55000, SDS-PAGE, x * 65000, SDS-PAGE | Alkalimonas collagenimarina |
| 65000 | - |
x * 55000, SDS-PAGE, x * 65000, SDS-PAGE | Alkalimonas collagenimarina |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Alkalimonas collagenimarina | D0VYQ9 | - |
- |
| Alkalimonas collagenimarina AC40T | D0VYQ9 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| by ammonium sulfate precipitation, to almost homogeneity, 364fold with recovery of 24.9% of the original activity. AcpII-C and AcpII-C-DELTAPA purified 109- and 219fold, with overall yields of 6.9% and 0.36%, respectively | Alkalimonas collagenimarina |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 2.2 | - |
toward keratin | Alkalimonas collagenimarina |
| 3 | 8 | toward casein | Alkalimonas collagenimarina |
| 10 | - |
toward elastin | Alkalimonas collagenimarina |
| 115.9 | - |
toward collagen | Alkalimonas collagenimarina |
| 143 | - |
toward gelatin | Alkalimonas collagenimarina |
Storage Stability
| Storage Stability | Organism |
|---|---|
| -20°C | Alkalimonas collagenimarina |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| casein + H2O | 38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein | Alkalimonas collagenimarina | ? | - |
? | |
| casein + H2O | 38% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on casein | Alkalimonas collagenimarina AC40T | ? | - |
? | |
| Collagen + H2O | 81% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on collagen | Alkalimonas collagenimarina | ? | - |
? | |
| Collagen + H2O | 81% activity relative to gelatin as substrate. Also purified AcpII-C and AcpII-C-DELTAPA act well on collagen | Alkalimonas collagenimarina AC40T | ? | - |
? | |
| Elastin + H2O | 7% activity relative to gelatin as substrate | Alkalimonas collagenimarina | ? | - |
? | |
| Gelatin + H2O | best substrate for AcpII. Also purified AcpII-C and AcpII-C-DELTAPA act well on gelatin | Alkalimonas collagenimarina | ? | - |
? | |
| Gelatin + H2O | best substrate for AcpII. Also purified AcpII-C and AcpII-C-DELTAPA act well on gelatin | Alkalimonas collagenimarina AC40T | ? | - |
? | |
| glutaryl-Ala-Ala-Pro-Leu-p-nitroanilide + H2O | relative hydrolysis rate is 22% | Alkalimonas collagenimarina | glutaryl-Ala-Ala-Pro-Leu + p-nitroaniline | - |
? | |
| Keratin + H2O | 1.5% activity relative to gelatin as substrate | Alkalimonas collagenimarina | ? | - |
? | |
| Keratin + H2O | 1.5% activity relative to gelatin as substrate | Alkalimonas collagenimarina AC40T | ? | - |
? | |
| methoxysuccinyl-Ala-Ile-Pro-Met-p-nitroanilide + H2O | is the most favorable substrate, relative hydrolysis rate is 100% | Alkalimonas collagenimarina | methoxysuccinyl-Ala-Ile-Pro-Met + p-nitroaniline | - |
? | |
| additional information | N-succinyl-Ala-Ala-Val-Ala-p-nitroanilide and keratin are poor substrates for AcpII, with relative hydrolysis rates of 1-1.5% of the activity toward N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide or gelatin, respectively | Alkalimonas collagenimarina | ? | - |
? | |
| additional information | N-succinyl-Ala-Ala-Val-Ala-p-nitroanilide and keratin are poor substrates for AcpII, with relative hydrolysis rates of 1-1.5% of the activity toward N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide or gelatin, respectively | Alkalimonas collagenimarina AC40T | ? | - |
? | |
| N-succinyl-Ala-Ala-Pro-Met-p-nitroanilide + H2O | relative hydrolysis rate is 39% | Alkalimonas collagenimarina | N-succinyl-Ala-Ala-Pro-Met + p-nitroaniline | - |
? | |
| N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O | relative hydrolysis rate is 94% | Alkalimonas collagenimarina | N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 55000, SDS-PAGE, x * 65000, SDS-PAGE | Alkalimonas collagenimarina |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AcpII | - |
Alkalimonas collagenimarina |
| Alkaline protease | - |
Alkalimonas collagenimarina |
| subtilisin-like protease | - |
Alkalimonas collagenimarina |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 45 | - |
at pH 9.5 in 100 mM glycine-NaOH buffer | Alkalimonas collagenimarina |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
residual activity is 80%, 60%, or 3% of the original activity after 15 min incubation at 45°C, 50°C, or 55°C, respectively. CaCl2 slightly increases the thermal stability of AcpII up to 45°C, and the residual activity is decreased to 80% after incubation at 50°C for 15 min | Alkalimonas collagenimarina |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.5 | 9 | in 100 mM glycine-NaOH buffer | Alkalimonas collagenimarina |
General Information
| General Information | Comment | Organism |
|---|---|---|
| physiological function | AcpII comprises a prepropeptide, a catalytic domain that includes a protease-associated domain, and tandem repeat prepeptidase C-terminal domains | Alkalimonas collagenimarina |
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Release 2023.1 (January 2023)
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